{"gene":"HRAS","run_date":"2026-06-10T01:55:22","timeline":{"discoveries":[{"year":1993,"finding":"A novel 37-kDa palmitoyl-protein thioesterase purified from bovine brain cytosol enzymatically removes palmitate from H-Ras. The thioesterase recognizes H-Ras only in its native conformation (bound to Mg2+ and guanine nucleotide), and it is sensitive to diethyl pyrocarbonate but insensitive to PMSF and NEM. Palmitoylated alpha subunits of heterotrimeric G proteins are also substrates.","method":"Enzyme purification to homogeneity, in vitro thioesterase assay with [3H]palmitate-labeled H-Ras produced in baculovirus, chemical inhibitor profiling","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — reconstituted in vitro enzymatic assay with purified enzyme and defined substrate, multiple inhibitor controls, single rigorous study","pmids":["7901201"],"is_preprint":false},{"year":1998,"finding":"H-Ras peptide and protein substrates bind protein farnesyltransferase (FTase) as an ionized thiolate: metal coordination of the substrate cysteine sulfur by the catalytic zinc ion lowers the pKa of the thiol (pKa ~6.3), generating a bound thiolate at physiological pH. The two substrates (H-Ras CAAX peptide and FPP) bind FTase synergistically, with affinity enhanced ~70-fold in the ternary complex.","method":"Fluorescence binding assays, pH-dependent affinity measurements, optical absorption spectroscopy of Co2+-substituted FTase, in vitro biochemical assay","journal":"Biochemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro reconstitution with purified enzyme, multiple orthogonal spectroscopic methods, mechanistic detail on catalytic zinc coordination","pmids":["9799520"],"is_preprint":false},{"year":1989,"finding":"Alternative splicing of the H-ras pre-mRNA normally suppresses p21 H-Ras expression: a negative-acting element in the last intron functions as an alternative exon; transcripts containing this exon are produced at low abundance (due to instability/defective processing) and are predicted to lack transforming potential. Mutations that prevent inclusion of this alternative exon increase H-ras expression and transforming efficiency.","method":"Gene reconstruction experiments, S1 nuclease analysis, mutational analysis of intronic elements, transcript identification","journal":"Cell","confidence":"High","confidence_rationale":"Tier 1 / Moderate — multiple orthogonal molecular biology methods (gene reconstruction, S1 analysis, mutational scanning) in a single rigorous study establishing the splicing control mechanism","pmids":["2667764"],"is_preprint":false},{"year":1995,"finding":"RHAMM (hyaluronan receptor) acts downstream of H-Ras in the transformation pathway: fibroblasts with a dominant suppressor mutant of RHAMM revert H-ras-transformed fibrosarcomas to a nontumorigenic, nonmetastatic phenotype, and antisense suppression of RHAMM renders fibroblasts resistant to ras transformation. Loss of functional RHAMM ablates focal adhesion kinase phosphorylation signaling within focal adhesions, preventing turnover in response to hyaluronan.","method":"Dominant suppressor transfection, antisense transfection, soft agar assays, in vivo tumorigenicity and metastasis assays, focal adhesion kinase phosphorylation analysis, genetic epistasis","journal":"Cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic epistasis (dominant suppressor + antisense) combined with defined biochemical readout (FAK phosphorylation) and in vivo tumor assays","pmids":["7541721"],"is_preprint":false},{"year":2002,"finding":"H-Ras signaling through the Raf/MEK/MAPK cascade requires endocytosis and endocytic recycling, whereas K-Ras signaling does not. Dominant-interfering dynamin-K44A selectively blocked H-Ras- but not K-Ras-mediated Raf-1 activation and PC12 differentiation. Wild-type Rab5 (stimulating endocytosis and recycling) potentiated H-Ras/Raf-1 signaling, while Rab5-Q79L (endocytosis without recycling) redistributed active H-Ras to enlarged endosomes and inhibited H-Ras/Raf-1 activation. H-Ras (but not K-Ras) signaling also required PI3K activity.","method":"Dominant-interfering dynamin expression, Rab5 mutant overexpression, Raf-1 kinase activity assay, subcellular fractionation/localization, PC12 differentiation assay","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple genetic tools (dominant-negative dynamin, activating and non-recycling Rab5 mutants) with isoform-specific readouts in two cell systems","pmids":["12077341"],"is_preprint":false},{"year":2007,"finding":"H-Ras enters cells via clathrin-independent endocytosis (CIE): it co-localizes with MHC class I CIE cargo and is sequestered in Arf6-Q67L vacuoles. Activated H-RasG12V induces macropinocytosis; incoming macropinosomes sequentially contain PIP2/PIP3, then PIP3/Rab5, then Rab5 alone. Arf6-Q67L traps H-Ras signaling in the PIP2/PIP3 stage, recruiting active ERK and Akt but not Rab5.","method":"Live fluorescence imaging, PH-domain reporters for phosphoinositides, Arf6 and Rab5 mutant expression, co-localization with CIE cargo markers","journal":"Molecular biology of the cell","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — live imaging with lipid reporters and genetic tools in a single lab; clear mechanistic staging of endosomal H-Ras signaling","pmids":["18094044"],"is_preprint":false},{"year":2009,"finding":"H-Ras protein is degraded via beta-TrCP-mediated polyubiquitylation and proteasomal degradation. H-Ras interacts with the WD40 domain of beta-TrCP; this ubiquitylation is stimulated by Axin or APC and inhibited by Wnt3a. In vivo, intravenous Wnt3a injection in mice reduces Ras levels in intestinal tissue, establishing Wnt/beta-catenin signaling as a regulator of H-Ras stability and providing a mechanistic basis for crosstalk between Wnt and Ras-ERK pathways.","method":"Co-immunoprecipitation, ubiquitylation assay, proteasome inhibitor experiments, Axin/APC/Wnt3a gain- and loss-of-function, in vivo Wnt3a injection in mice","journal":"Journal of cell science","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP mapping to WD40 domain, ubiquitylation assay, multiple gain/loss-of-function reagents, in vivo validation","pmids":["19240121"],"is_preprint":false},{"year":2011,"finding":"Metabolic stress (high palmitate/glucose) oxidizes H-Ras cysteine thiols at Cys181/184, detected by MALDI-TOF/TOF mass spectrometry, which prevents palmitoylation. Loss of palmitoylation shifts H-Ras from plasma membrane to Golgi, decreases growth factor-stimulated ERK phosphorylation (~84% reduction), and increases apoptotic signaling; these effects are prevented by wild-type but not C181/184S H-Ras, establishing oxidation of palmitoylation-site cysteines as the causal mechanism.","method":"MALDI-TOF/TOF mass spectrometry of Cys181/184, palmitoylation assay, subcellular fractionation, ERK phosphorylation measurement, apoptosis assay, WT vs. C181/184S mutant rescue, MnSOD overexpression","journal":"FASEB journal","confidence":"High","confidence_rationale":"Tier 1 / Strong — direct mass spectrometric identification of oxidized cysteines, site-specific mutagenesis (C181/184S), and multiple orthogonal functional readouts in a single study","pmids":["22085642"],"is_preprint":false},{"year":2014,"finding":"H-Ras forms dimers on membrane surfaces via a protein-protein interface involving the switch II region (near the nucleotide binding cleft). A Y64A point mutation in switch II abolishes dimer formation. Dimerization requires the membrane surface and does not occur in solution at comparable densities; it does not require lipid anchor clustering. The 2D dimerization Kd is ~1 × 10^3 molecules/μm^2, and no higher-order oligomers are observed.","method":"Fluorescence correlation spectroscopy, photon counting histogram analysis, time-resolved fluorescence anisotropy, single-molecule tracking, step photobleaching, supported lipid bilayer reconstitution, site-directed mutagenesis (Y64A)","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1 / Strong — multiple orthogonal single-molecule and ensemble biophysical methods on reconstituted membranes, mutagenesis validation, quantitative Kd measurement","pmids":["24516166"],"is_preprint":false},{"year":2013,"finding":"HRas GTPase, activated downstream of EGFR signaling, is required for HCV entry into hepatocytes. Proteomic analysis showed HRas associates with tetraspanin CD81, claudin-1, integrin β1, and Rap2B in hepatocyte membranes. HRas signaling mediates lateral membrane diffusion of CD81, enabling tetraspanin receptor complex assembly necessary for HCV entry.","method":"Proteomic/MS analysis of HRas-associated complex, RNAi knockdown of HRas, lateral membrane diffusion assays, HCV infection assay, Co-IP","journal":"Cell host & microbe","confidence":"High","confidence_rationale":"Tier 2 / Strong — MS-based interactome identification, RNAi functional validation, lateral diffusion measurement, and infection assay provide multiple orthogonal lines of evidence","pmids":["23498955"],"is_preprint":false},{"year":2004,"finding":"In H-Ras-transformed cells, elevated MAPK activity (from H-Ras) transcriptionally induces Egr1, which in turn upregulates KLF5; KLF5 then activates cyclin D1 expression. MEK inhibitors reduce KLF5 and abolish anchorage-independent growth. siRNA knockdown of KLF5 in H-Ras-transformed cells reduces proliferation and colony formation, establishing KLF5 as a downstream mediator of H-Ras oncogenic transformation via the MEK/ERK → Egr1 → KLF5 → cyclin D1 axis.","method":"MEK inhibitor treatment, siRNA knockdown of KLF5 and Egr1, soft agar colony formation, proliferation assays, qRT-PCR, Western blot","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — RNAi epistasis at two nodes (Egr1, KLF5) with defined phenotypic readouts; single lab","pmids":["15077182"],"is_preprint":false},{"year":2016,"finding":"H-ras inhibits the Hippo tumor suppressor pathway by promoting formation of inactive Mst1/Mst2 heterodimers via an ERK-dependent mechanism. Mst1/Mst2 heterodimers (mediated by SARAH domains) have much lower kinase activity than homodimers. Cells lacking Mst1 (unable to form heterodimers) are resistant to H-ras-mediated transformation and maintain active Hippo signaling.","method":"Co-immunoprecipitation of Mst1/Mst2 heterodimers, in vitro kinase assay comparing homo- vs. heterodimer activity, MEK inhibitor epistasis, Mst1 knockout cells, focus formation assay","journal":"Current biology : CB","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — Co-IP demonstrating heterodimer formation, in vitro kinase activity comparison, genetic KO epistasis, and transformation assay provide multiple orthogonal mechanistic validations","pmids":["27238285"],"is_preprint":false},{"year":2017,"finding":"Aurora kinase A (Aurora A) physically interacts with H-Ras through its kinase domain binding the N-terminal domain of H-Ras; Aurora A, H-Ras, and Raf-1 exist in a trimeric complex. Aurora A enhances H-Ras binding to Raf-1 and potentiates H-Ras-mediated MAPK signaling in an active H-Ras-dependent manner.","method":"Co-immunoprecipitation, domain-mapping pulldown assays, MAPK signaling readouts, overexpression of Aurora A with active/inactive H-Ras","journal":"Oncotarget","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal Co-IP with domain mapping and functional MAPK readout; single lab","pmids":["28177880"],"is_preprint":false},{"year":2013,"finding":"H-Ras transfers between cells via tunneling nanotubes (TNTs) connecting B and T cells. GFP-H-Ras diffuses freely in the TNT membrane (FRAP), and PM patches enriched in GFP-H-Ras and CD86 transfer to T-cell surfaces. Transfer is dependent on normal post-translational lipidation (palmitoylation/farnesylation) of H-Ras for plasma membrane anchorage.","method":"Optical tweezers cell trapping, 4D spinning-disk confocal live imaging, FRAP on TNT membranes, FACS, lipidation-defective mutant H-Ras controls","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — live imaging and FRAP with lipidation-mutant controls; single lab, single study","pmids":["23868059"],"is_preprint":false},{"year":2008,"finding":"The HRAS p.Lys117Arg mutation (Costello syndrome) constitutively activates the RAS/MAPK pathway through a mechanism distinct from codon 12/13 mutations: it increases the nucleotide dissociation rate ~80-fold while retaining normal intrinsic GTP hydrolysis and GAP responsiveness. The crystal structure of HRAS p.Lys117Arg shows an altered side-chain interaction pattern that unfavorably affects nucleotide binding.","method":"Fluorescence nucleotide exchange/dissociation assays, GTPase activity assay, GAP-stimulated hydrolysis assay, X-ray crystallography of recombinant mutant protein","journal":"Human mutation","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structure plus multiple in vitro biochemical assays (exchange rate, hydrolysis, GAP responsiveness) on recombinant mutant protein","pmids":["17979197"],"is_preprint":false},{"year":2016,"finding":"TRPML1 (encoded by MCOLN1) maintains oncogenic HRAS in signaling-competent nanoclusters at the plasma membrane by mediating cholesterol de-esterification and transport. TRPML1 inhibition disrupts cholesterol distribution, attenuates HRAS nanoclustering and plasma membrane abundance, and reduces ERK phosphorylation and proliferation selectively in HRAS-mutant cancer cells.","method":"MCOLN1 knockdown and pharmacological TRPML1 inhibition, cholesterol distribution assays, HRAS nanoclustering quantification, ERK phosphorylation, proliferation assays in HRAS-mutant vs. wild-type cell lines","journal":"EMBO reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic KD and pharmacological inhibition with mechanistic cholesterol/nanoclustering readouts; single lab","pmids":["30787043"],"is_preprint":false},{"year":2016,"finding":"Endogenous H-Ras (mVenus-HRas knock-in) is primarily located at the plasma membrane and in small amounts in tubular recycling endosomes; EGF stimulation causes fast but transient HRas activation. EGFR endocytosis physically separates EGFR-Grb2 complexes from HRas, terminating Ras-mediated signaling, while sustained minimal ERK activation is maintained by a small pool of active EGFRs remaining at the plasma membrane.","method":"CRISPR/gene-editing to tag endogenous HRas with mVenus, live fluorescence imaging, subcellular colocalization, EGFR/Grb2 endosomal tracking, surface biotinylation, selective inactivation of surface EGFRs, ERK/MEK phosphorylation time courses","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — endogenous tagging via gene editing, multiple imaging and biochemical readouts, and specific surface EGFR inactivation experiments in a single rigorous study","pmids":["26858456"],"is_preprint":false},{"year":2019,"finding":"Compartment-specific HRAS signaling: HRAS signaling is strongest from the cell membrane (highest kinase activation) but regulates the largest number of genes from the endoplasmic reticulum. Signaling from the Golgi apparatus activates TP53-dependent cell survival. Different subcellular locations generate distinct protein interaction profiles and kinase activation patterns that differentially regulate gene transcription.","method":"Compartment-targeted HRAS constructs, phosphoproteomics, protein interaction proteomics, transcriptomics, integrated network analysis","journal":"Cell reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple omics methods with compartment-targeted constructs; single lab, complex integrated analysis","pmids":["30865897"],"is_preprint":false},{"year":2019,"finding":"QPCT binds directly to HRAS and attenuates its ubiquitination, thereby increasing HRAS protein stability and activating the ERK pathway. This interaction was identified by human proteome microarray and validated by co-IP and confocal colocalization. QPCT upregulation (driven by reduced promoter methylation) confers sunitinib resistance in renal cell carcinoma via HRAS stabilization.","method":"Human proteome microarray, co-immunoprecipitation, confocal colocalization, ubiquitination assay, siRNA knockdown, overexpression, ERK pathway readouts","journal":"Theranostics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — proteome microarray binding identification confirmed by Co-IP and colocalization, with mechanistic ubiquitination assay; single lab","pmids":["31534544"],"is_preprint":false},{"year":2023,"finding":"m6A modification at three specific sites in the HRAS 3' UTR (but not KRAS or NRAS 3' UTR) promotes H-Ras protein expression by enhancing translational elongation. This modification is regulated by the m6A eraser FTO (which removes the modification) and read by YTHDF1 (which promotes translation); YTHDF2 and YTHDF3 are not involved. Targeting HRAS m6A modification decreases cancer cell proliferation and metastasis.","method":"m6A sequencing, luciferase reporter assays with mutated m6A sites, YTHDF1/2/3 and FTO knockdown/overexpression, polysome profiling for translational elongation, proliferation and metastasis assays","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple writer/reader/eraser perturbation experiments with specific 3' UTR site mapping; single lab","pmids":["36996116"],"is_preprint":false},{"year":2011,"finding":"The HRAS promoter contains two G-rich elements (hras-1 and hras-2) that fold into G-quadruplex structures acting as transcriptional repressors. MAZ binds both quadruplexes; Sp1 binds only qhras-1. hnRNP A1 binds the complementary C-rich i-motif structures and unfolds them (demonstrated by FRET and CD), and is required for HRAS transcriptional activation (knockdown reduces transcript to ~44%).","method":"Chromatin immunoprecipitation (ChIP), EMSA, promoter mutagenesis (blocking or stabilizing quadruplexes), transcription assays, quadruplex-stabilizing ligands, RNAi knockdown","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP in cells combined with in vitro EMSA and mutagenesis; multiple regulatory elements characterized; single lab","pmids":["21931711"],"is_preprint":false},{"year":2015,"finding":"hnRNP A1 binds the C-rich i-motif structures in the HRAS promoter GC-elements (hras-1 and hras-2), unfolds them upon binding (demonstrated by FRET and CD), and is necessary for HRAS transcriptional activation. Knockdown of hnRNP A1 reduces HRAS transcript to 44% of control. Sequestration of i-motif-binding proteins by decoy oligonucleotides significantly inhibits HRAS transcription.","method":"ChIP, EMSA, FRET, CD spectroscopy, shRNA knockdown of hnRNP A1, decoy oligonucleotide competition","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — biophysical (FRET, CD) and cell-based (ChIP, shRNA) methods concordant; single lab replicating and extending prior findings","pmids":["26674223"],"is_preprint":false},{"year":2023,"finding":"The crystal structure of the HRAS oncogene promoter i-motif (iHRAS) was solved at 1.77 Å resolution. iHRAS folds into a double hairpin forming an antiparallel i-motif dimer capped by loops; six C-C+ base pairs form each i-motif core, extended by a G-G base pair and cytosine stacking. Extensive canonical and non-canonical base pairing stabilizes the connecting region and loops.","method":"X-ray crystallography at 1.77 Å resolution","journal":"Angewandte Chemie (International ed. in English)","confidence":"High","confidence_rationale":"Tier 1 / Strong — atomic resolution crystal structure with full structural determination","pmids":["36995904"],"is_preprint":false},{"year":2010,"finding":"Endoglin (TGF-β co-receptor) inhibits H-Ras expression and its oncogenic transformation capacity. TGF-β1 increases H-Ras mRNA and protein via ALK5 and the Ras-MAPK pathway (Smad4-independent); endoglin attenuates this TGF-β1-mediated induction. Endoglin also inhibits Ras-GTP levels, phospho-MEK, and phospho-ERK in H-Ras oncogene-expressing cells, and suppresses transformation by H-RasQ61K and H-RasG12V in NIH3T3 focus formation assays.","method":"H-Ras promoter transactivation assay, Northern/Western blot, Ras-GTP pulldown, MEK/ERK phosphorylation, NIH3T3 focus formation assay, ALK5 inhibitor epistasis","journal":"Carcinogenesis","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple biochemical readouts and functional transformation assay; single lab","pmids":["20884686"],"is_preprint":false},{"year":2014,"finding":"CIP2A physically associates with H-Ras, leading to activation of the MEK/ERK signaling pathway and promotion of epithelial-mesenchymal transition (EMT) in cervical cancer cells. The interaction was identified by pulldown/MS and validated by bilateral co-immunoprecipitation.","method":"Pulldown assay, mass spectrometric peptide sequencing, bilateral co-immunoprecipitation, MEK/ERK phosphorylation assay, EMT marker analysis","journal":"Cancer letters","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — interaction identified by MS and confirmed by reciprocal Co-IP with functional downstream readout; single lab","pmids":["25458953"],"is_preprint":false},{"year":2022,"finding":"HRAS is uniquely dependent on farnesyltransferase (FTase) for prenylation and membrane localization; NRAS and KRAS can use geranylgeranyl transferase as a bypass. Tipifarnib (FTase inhibitor) reduces HRAS processing and plasma membrane localization, decreasing GTP-bound HRAS and downstream effector pathway signaling (RAS effector pathways). This selectively inhibits growth of HRAS-mutant but not NRAS- or KRAS-mutant RMS xenografts in vivo.","method":"HRAS prenylation/processing assays, plasma membrane fractionation, GTP-bound HRAS (active Ras) pulldown, RAS effector pathway signaling (Western blot), 2D/3D cell growth assays, in vivo xenograft models with genomic RAS mutation stratification","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 2 / Strong — direct biochemical demonstration of FTase-dependent HRAS processing and membrane localization with mutation-stratified in vivo validation; multiple orthogonal methods","pmids":["35459782"],"is_preprint":false},{"year":2022,"finding":"Oncogenic HRAS mutations (G12S, G12A) inhibit AMPK signaling, leading to impaired mitochondrial proteostasis and defective oxidative phosphorylation. This was demonstrated in CS mouse heart/skeletal muscle, patient fibroblasts, hiPSC-derived cardiomyocytes, and zebrafish models. Pharmacological activation of mitochondrial bioenergetics restored organelle function and reduced cardiac hypertrophy.","method":"CS mouse model (HrasG12S), patient skin fibroblasts, hiPSC-derived cardiomyocytes, HrasG12V zebrafish model, lentiviral HRAS mutant constructs, AMPK pathway analysis, mitochondrial function assays (respiration, proteostasis), pharmacological rescue","journal":"The Journal of clinical investigation","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple independent model systems (mouse, human cells, fish, iPSC), convergent pathway (AMPK) identification, and pharmacological rescue; well-controlled study","pmids":["35230976"],"is_preprint":false},{"year":2021,"finding":"Inhibition of MEK/MAPK pathway signaling with a MEK inhibitor rescues HrasG12V-induced skeletal myopathy in a Costello syndrome mouse model. Activated HrasG12V causes skeletal myopathy via inhibition of embryonic myogenesis and myofiber formation, involving hyperactivation of Ras/MAPK and PI3K/AKT pathways and significant reduction in p38 signaling. MEK inhibition rescues the myopathy both in vitro and in vivo.","method":"HrasG12V knock-in mouse model, muscle histomorphometry, in vitro myogenesis assays, MEK inhibitor treatment in vitro and in vivo, phosphoprotein analysis (MAPK, AKT, p38), global transcriptomics","journal":"Disease models & mechanisms","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic mouse model with pharmacological rescue and multiple signaling readouts; single lab","pmids":["34553752"],"is_preprint":false},{"year":2023,"finding":"Mutant HRAS drives metastasis of head and neck cancer by suppressing the Hippo pathway and stabilizing YAP1, which transcriptionally activates AXL. Farnesyltransferase inhibitor tipifarnib activates the Hippo pathway, reduces nuclear YAP1 export, suppresses YAP1-mediated AXL expression, attenuates VEGFA/VEGFC expression, and reduces lymphovascular angiogenesis and metastasis. AXL depletion phenocopies tipifarnib in blocking invasion and metastasis.","method":"HRASmut human HNC cell lines, patient-derived xenografts, syngeneic mouse model, genetic (siRNA/shRNA) and pharmacological (tipifarnib) manipulation, targeted proteomics, Hippo pathway signaling analysis, in vivo metastasis assays","journal":"Cancer research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo PDX and syngeneic models with genetic and pharmacological validation; single lab","pmids":["36753744"],"is_preprint":false},{"year":1987,"finding":"Oncogenic forms of H-ras (but not proto-oncogenic forms) completely suppress skeletal myoblast differentiation (fusion, nicotinic acetylcholine receptor and creatine kinase induction) at the level of muscle-specific mRNA accumulation, independent of effects on cell proliferation. This demonstrates H-ras mimics the differentiation-inhibitory effects of FGF and TGF-beta.","method":"DNA-mediated gene transfer into C2 myoblasts, myotube fusion assay, muscle-specific gene expression (Northern blot for AChR and creatine kinase mRNA), growth curve analysis","journal":"Molecular and cellular biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — gain-of-function with oncogenic vs. proto-oncogenic forms, multiple differentiation readouts; foundational study, single lab","pmids":["3600660"],"is_preprint":false},{"year":2016,"finding":"SPRED1 specifically perturbs K-ras (but not H-ras) membrane organization and ERK signaling at the plasma membrane by becoming enriched in acidic membrane domains. However, SPRED1 does block the positive effects of galectin-1 on H-ras nanoclustering. Legius syndrome-associated SPRED1 mutations show diminished binding to both galectin-1 and B-Raf.","method":"SPRED1 overexpression and membrane domain analysis, H-ras vs. K-ras ERK signaling assays, galectin-1 co-expression, Legius syndrome mutant analysis, plasma membrane fractionation","journal":"Molecular and cellular biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct comparison of H-ras vs. K-ras with mechanistic membrane domain analysis and disease mutant validation; single lab","pmids":["27503857"],"is_preprint":false},{"year":2004,"finding":"H-Ras and K-Ras exert distinct effects on apoptosis in endometrial cells: activated H-Ras rescues cells from apoptosis (dependent on PI3K/Akt), whereas activated K-Ras promotes apoptosis (dependent on Raf/MEK/MAPK). These isoform-specific differences depend on effector domain interactions, as demonstrated by K-Ras effector domain mutants (K12V35S preferentially activating MAPK/apoptosis; K12V40C preferentially activating Akt/survival).","method":"Stable expression of activated H-Ras and K-Ras, MEK inhibitor (U0126) and PI3K inhibitor (LY294002) epistasis, K-Ras effector domain mutants (35S and 40C), apoptosis assays, MAPK/Akt phosphorylation","journal":"Cancer research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — effector domain mutant epistasis with isoform-specific inhibitor experiments; single lab","pmids":["15087391"],"is_preprint":false}],"current_model":"HRAS is a lipid-anchored small GTPase that cycles between GDP-bound (inactive) and GTP-bound (active) states, undergoes sequential post-translational modifications (farnesylation by FTase — uniquely obligate for HRAS unlike KRAS/NRAS — followed by proteolysis, methylation, and palmitoylation at Cys181/184), and signals from distinct plasma membrane nanoclusters, endosomes, Golgi, and ER compartments to activate downstream cascades including Raf/MEK/ERK and PI3K/Akt; its membrane localization, palmitoylation-cycle dynamics, and signaling output are regulated by redox modification of palmitoylation-site cysteines, beta-TrCP/Wnt-mediated ubiquitin-proteasomal degradation, QPCT-mediated stabilization, m6A epitranscriptomic control of translation, G-quadruplex/i-motif promoter structures (bound by MAZ, Sp1, and hnRNP A1), ERK-dependent promotion of Mst1/Mst2 heterodimer formation to suppress the Hippo pathway, Aurora A-scaffold enhancement of Raf-1 recruitment, and TRPML1-mediated cholesterol transport that maintains plasma membrane nanoclustering; oncogenic HRAS mutations additionally impair AMPK-mitochondrial homeostasis and drive metastasis via YAP1-AXL axis activation."},"narrative":{"mechanistic_narrative":"HRAS is a lipid-anchored small GTPase that signals through the Raf/MEK/ERK and PI3K/Akt cascades to drive proliferation, transformation, and developmental signaling [PMID:15077182, PMID:15087391]. Its activity depends on a sequence of post-translational membrane-targeting events: FTase catalyzes farnesylation through a mechanism in which the catalytic zinc coordinates the CAAX cysteine as an ionized thiolate, and FTase substrate is bound synergistically with farnesyl pyrophosphate [PMID:9799520]; HRAS is uniquely obligate for FTase, since unlike NRAS and KRAS it cannot bypass FTase via geranylgeranyltransferase, making HRAS-mutant tumors selectively sensitive to FTase inhibition [PMID:35459782]. A dynamic palmitoylation cycle at Cys181/184 — installed by palmitoylation and reversed by a palmitoyl-protein thioesterase that recognizes only native, nucleotide-bound H-Ras [PMID:7901201] — governs membrane distribution; oxidation of these cysteines under metabolic stress blocks palmitoylation, redistributes HRAS from plasma membrane to Golgi, and suppresses ERK signaling [PMID:22085642]. HRAS signaling is spatially partitioned: it activates strongest from the plasma membrane, regulates the broadest transcriptional program from the ER, and engages TP53-dependent survival from the Golgi [PMID:30865897], with plasma-membrane signaling requiring nanocluster organization maintained by TRPML1-dependent cholesterol transport [PMID:30787043] and HRAS-specific endocytic recycling for Raf-1 activation [PMID:12077341, PMID:26858456]. On the membrane HRAS forms switch-II-mediated dimers abolished by the Y64A mutation [PMID:24516166]. Downstream, HRAS-ERK signaling drives transformation through an Egr1→KLF5→cyclin D1 axis [PMID:15077182], suppresses the Hippo pathway by promoting low-activity Mst1/Mst2 heterodimers [PMID:27238285], and in mutant head-and-neck cancer stabilizes YAP1 to transcriptionally activate AXL and drive metastasis [PMID:36753744]. HRAS protein abundance is set by beta-TrCP/Wnt-regulated ubiquitin-proteasomal degradation [PMID:19240121], QPCT-mediated stabilization [PMID:31534544], alternative-splicing suppression of p21 expression [PMID:2667764], and m6A-dependent translational control of the HRAS 3' UTR via FTO and YTHDF1 [PMID:36996116]. Germline activating HRAS mutations cause Costello syndrome: the p.Lys117Arg mutation accelerates nucleotide dissociation ~80-fold while retaining normal hydrolysis [PMID:17979197], and oncogenic G12 mutations produce skeletal myopathy and cardiac hypertrophy through MEK/MAPK hyperactivation and AMPK-mitochondrial dysfunction [PMID:35230976, PMID:34553752].","teleology":[{"year":1987,"claim":"Established that oncogenic HRAS does more than drive proliferation — it actively blocks a differentiation program, defining HRAS as a determinant of cell fate.","evidence":"Gene transfer of oncogenic vs proto-oncogenic H-ras into C2 myoblasts with muscle-specific gene expression readouts","pmids":["3600660"],"confidence":"Medium","gaps":["Molecular intermediates linking HRAS to suppression of muscle-specific transcription not defined","Did not distinguish effector pathways responsible"]},{"year":1989,"claim":"Showed that HRAS protein levels are constrained pre-translationally, identifying alternative splicing of an intronic exon as a negative control over p21 H-Ras expression and transforming potential.","evidence":"Gene reconstruction, S1 nuclease analysis, and intronic mutational scanning","pmids":["2667764"],"confidence":"High","gaps":["Trans-acting splicing factors not identified","Physiological signals that modulate exon inclusion unknown"]},{"year":1993,"claim":"Identified an enzyme that reverses HRAS palmitoylation, establishing that the palmitoylation state is dynamically regulated and conformation-dependent.","evidence":"Purification of a 37-kDa palmitoyl-protein thioesterase from bovine brain and in vitro assay on native, nucleotide-bound H-Ras","pmids":["7901201"],"confidence":"High","gaps":["In vivo relevance to HRAS membrane cycling not tested","Gene identity of the thioesterase not established here"]},{"year":1995,"claim":"Placed RHAMM-dependent focal adhesion signaling downstream of HRAS, connecting HRAS transformation to adhesion turnover and metastasis.","evidence":"Dominant suppressor and antisense RHAMM in H-ras-transformed fibroblasts with tumorigenicity, metastasis, and FAK phosphorylation readouts","pmids":["7541721"],"confidence":"High","gaps":["Direct biochemical link between HRAS and RHAMM not shown","Mechanism of FAK regulation downstream of RHAMM unresolved"]},{"year":1998,"claim":"Defined the catalytic chemistry of HRAS prenylation, showing the CAAX cysteine binds FTase as a zinc-coordinated thiolate with synergistic FPP binding.","evidence":"Fluorescence binding, pH-dependent affinity, and Co2+-substituted FTase spectroscopy","pmids":["9799520"],"confidence":"High","gaps":["Does not address downstream proteolysis/methylation steps","In-cell kinetics not measured"]},{"year":2002,"claim":"Revealed isoform-specific spatial requirements, showing HRAS (unlike KRAS) requires endocytosis/recycling and PI3K for Raf-1 activation.","evidence":"Dominant-negative dynamin and Rab5 mutants with Raf-1 kinase and PC12 differentiation assays","pmids":["12077341"],"confidence":"High","gaps":["Identity of the signaling-competent endosomal compartment not fully resolved","Mechanism linking recycling to Raf-1 activation unknown"]},{"year":2004,"claim":"Connected HRAS-ERK output to a transcriptional transformation program and to isoform-specific apoptotic control via effector selection.","evidence":"siRNA epistasis at Egr1/KLF5 and effector-domain mutant analysis with inhibitor epistasis in two cell systems","pmids":["15077182","15087391"],"confidence":"Medium","gaps":["Generality of Egr1→KLF5→cyclin D1 axis across tissues untested","Structural basis of effector-domain preference not defined"]},{"year":2007,"claim":"Mapped active HRAS through stages of clathrin-independent endocytosis and macropinocytosis, defining where compartmentalized ERK/Akt activation occurs.","evidence":"Live imaging with phosphoinositide reporters and Arf6/Rab5 mutants","pmids":["18094044"],"confidence":"Medium","gaps":["Single-lab imaging study","Functional consequences of each endosomal stage on transcription not assessed"]},{"year":2008,"claim":"Showed a Costello syndrome mutation activates HRAS by a novel biochemical route — accelerated nucleotide dissociation — distinct from canonical hydrolysis-defective G12/G13 mutations.","evidence":"Nucleotide exchange/hydrolysis/GAP assays plus crystal structure of recombinant p.Lys117Arg","pmids":["17979197"],"confidence":"High","gaps":["Cellular signaling magnitude relative to G12 mutants not quantified here","GEF dependence of the fast-cycling mutant unaddressed"]},{"year":2009,"claim":"Defined ubiquitin-proteasomal degradation of HRAS by beta-TrCP and showed Wnt/Axin/APC signaling sets HRAS abundance, establishing Wnt-Ras crosstalk.","evidence":"Reciprocal Co-IP to the beta-TrCP WD40 domain, ubiquitylation assays, gain/loss-of-function, and in vivo Wnt3a injection","pmids":["19240121"],"confidence":"High","gaps":["Phosphodegron on HRAS not mapped","Whether oncogenic mutants escape this degradation untested"]},{"year":2010,"claim":"Identified endoglin and TGF-beta/ALK5 as transcriptional and signaling regulators of HRAS expression and transformation capacity.","evidence":"Promoter transactivation, Ras-GTP pulldown, MEK/ERK readouts, and NIH3T3 focus formation with ALK5 inhibitor epistasis","pmids":["20884686"],"confidence":"Medium","gaps":["Direct cis-elements mediating TGF-beta induction not mapped","Single-lab study"]},{"year":2011,"claim":"Connected metabolic redox state to HRAS localization by showing oxidation of palmitoylation-site cysteines blocks palmitoylation and reroutes HRAS off the plasma membrane.","evidence":"MALDI-TOF/TOF identification of oxidized Cys181/184, C181/184S rescue, fractionation, and ERK/apoptosis readouts","pmids":["22085642"],"confidence":"High","gaps":["Reversibility of oxidation in vivo not quantified","Enzymes mediating re-reduction unidentified"]},{"year":2011,"claim":"Defined a structural/transcriptional control layer at the HRAS promoter, with G-quadruplex repressors bound by MAZ/Sp1 and i-motifs unfolded by hnRNP A1 to activate transcription.","evidence":"ChIP, EMSA, promoter mutagenesis, FRET/CD, and RNAi in cells","pmids":["21931711","26674223"],"confidence":"Medium","gaps":["In vivo dynamics of quadruplex/i-motif folding not measured","Quantitative contribution to endogenous HRAS expression incomplete"]},{"year":2013,"claim":"Extended HRAS biology to intercellular and pathogen contexts, showing HRAS transfers between cells via tunneling nanotubes and is required for HCV receptor complex assembly.","evidence":"Optical-tweezer/FRAP imaging with lipidation mutants; proteomics, RNAi, and lateral diffusion plus HCV infection assays","pmids":["23868059","23498955"],"confidence":"Medium","gaps":["Physiological frequency and significance of TNT transfer unknown","How HRAS GTPase activity drives CD81 lateral diffusion unresolved"]},{"year":2014,"claim":"Demonstrated HRAS dimerizes on membranes through a switch-II interface, defining a membrane-dependent oligomeric state relevant to effector activation.","evidence":"FCS, photon-counting histogram, single-molecule tracking, supported bilayer reconstitution, and Y64A mutagenesis","pmids":["24516166"],"confidence":"High","gaps":["Functional consequence of dimerization for Raf activation not directly tested","Role of dimer interface in cells unconfirmed"]},{"year":2014,"claim":"Identified CIP2A as a direct HRAS partner driving MEK/ERK activation and EMT, expanding the HRAS interactome in cancer.","evidence":"Pulldown/MS and bilateral Co-IP with EMT and signaling readouts","pmids":["25458953"],"confidence":"Medium","gaps":["Interaction interface not mapped","Single cancer-cell context"]},{"year":2016,"claim":"Resolved how HRAS signaling is terminated and sustained, showing endogenous HRAS is plasma-membrane/recycling-endosome localized and that EGFR endocytosis spatially separates EGFR-Grb2 from HRAS.","evidence":"CRISPR mVenus-HRas knock-in, live imaging, surface EGFR inactivation, and ERK/MEK time courses","pmids":["26858456"],"confidence":"High","gaps":["Generality across receptor systems untested","Quantitative contribution of recycling-endosome pool to signaling unresolved"]},{"year":2016,"claim":"Linked HRAS to Hippo suppression and defined membrane-organization regulators, showing ERK promotes inactive Mst1/Mst2 heterodimers and that TRPML1 cholesterol transport maintains HRAS nanoclusters.","evidence":"Co-IP and in vitro kinase comparison with Mst1 KO; MCOLN1 knockdown/inhibition with nanoclustering and ERK readouts","pmids":["27238285","30787043"],"confidence":"Medium","gaps":["Direct biochemical step from ERK to heterodimer formation incompletely defined","How cholesterol geometry enforces nanoclustering not structurally resolved"]},{"year":2016,"claim":"Clarified isoform-specific membrane regulation by SPRED1/galectin-1, distinguishing HRAS from KRAS nanocluster control.","evidence":"SPRED1 membrane-domain analysis, galectin-1 co-expression, and Legius syndrome mutant binding","pmids":["27503857"],"confidence":"Medium","gaps":["Mechanism by which SPRED1 blocks galectin-1's positive effect on HRAS unresolved","Single-lab study"]},{"year":2017,"claim":"Identified Aurora A as a scaffolding partner enhancing HRAS-Raf-1 binding within a trimeric complex, adding a kinase-independent layer to HRAS-MAPK activation.","evidence":"Reciprocal Co-IP, domain-mapping pulldowns, and MAPK readouts with active/inactive HRAS","pmids":["28177880"],"confidence":"Medium","gaps":["Structural basis of the trimeric complex unknown","In vivo relevance untested"]},{"year":2019,"claim":"Established that HRAS signaling identity is location-encoded, with distinct kinase, interaction, and transcriptional outputs from plasma membrane, ER, and Golgi.","evidence":"Compartment-targeted HRAS constructs with integrated phospho/interaction/transcriptomics","pmids":["30865897"],"confidence":"Medium","gaps":["Endogenous compartmental flux not measured","Targeted constructs may not recapitulate native localization stoichiometry"]},{"year":2019,"claim":"Identified QPCT as a direct HRAS-stabilizing partner that attenuates ubiquitination and confers drug resistance, complementing the beta-TrCP degradation axis.","evidence":"Proteome microarray, Co-IP, colocalization, ubiquitination assay, and ERK readouts in renal cell carcinoma","pmids":["31534544"],"confidence":"Medium","gaps":["Whether QPCT competes with beta-TrCP directly unknown","Interaction interface not mapped"]},{"year":2022,"claim":"Defined the unique FTase-dependence of HRAS and validated FTase inhibition as a mutation-selective therapeutic strategy.","evidence":"Processing, membrane fractionation, active-HRAS pulldown, and mutation-stratified RMS xenografts with tipifarnib","pmids":["35459782"],"confidence":"High","gaps":["Resistance mechanisms to sustained FTase inhibition not addressed","Effect on non-canonical HRAS compartments untested"]},{"year":2022,"claim":"Linked oncogenic HRAS to metabolic/organismal pathology, showing G12 mutations impair AMPK signaling and mitochondrial proteostasis in Costello syndrome models.","evidence":"CS mouse, patient fibroblasts, hiPSC cardiomyocytes, and zebrafish with AMPK and mitochondrial assays and pharmacological rescue","pmids":["35230976"],"confidence":"High","gaps":["Direct biochemical link from HRAS to AMPK inhibition not defined","Whether ERK pathway mediates the mitochondrial defect untested"]},{"year":2023,"claim":"Established m6A epitranscriptomic control specific to the HRAS 3' UTR, where FTO and YTHDF1 regulate translational elongation and tumor phenotypes.","evidence":"m6A-seq, site-mutated luciferase reporters, FTO/YTHDF perturbations, and polysome profiling","pmids":["36996116"],"confidence":"Medium","gaps":["Upstream signals controlling HRAS 3' UTR methylation unknown","Single-lab study"]},{"year":2023,"claim":"Resolved the atomic structure of the HRAS promoter i-motif and defined a YAP1-AXL metastatic axis driven by mutant HRAS, deepening both transcriptional-control and oncogenic-output models.","evidence":"1.77 Å crystal structure of iHRAS; HRASmut HNC cell/PDX/syngeneic models with tipifarnib and AXL depletion","pmids":["36995904","36753744"],"confidence":"Medium","gaps":["In-cell relevance of the crystallographic i-motif fold not confirmed","Mechanism of YAP1 stabilization by mutant HRAS incompletely defined"]},{"year":null,"claim":"How the multiple layers of HRAS abundance control (splicing, beta-TrCP degradation, QPCT stabilization, m6A translation, promoter quadruplex/i-motif structures) are integrated and coordinated with compartment-specific signaling in normal vs. oncogenic cells remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unified model linking expression-level control to spatial signaling output","Crosstalk among the regulatory layers untested in a single system"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0003924","term_label":"GTPase activity","supporting_discovery_ids":[14,25,9]},{"term_id":"GO:0060089","term_label":"molecular transducer activity","supporting_discovery_ids":[10,17,31]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[7,16,15,8]},{"term_id":"GO:0005768","term_label":"endosome","supporting_discovery_ids":[4,5,16]},{"term_id":"GO:0005794","term_label":"Golgi apparatus","supporting_discovery_ids":[7,17]},{"term_id":"GO:0005783","term_label":"endoplasmic reticulum","supporting_discovery_ids":[17]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[4,10,11,17]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[14,26,27,28]},{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[1,6,18,25]}],"complexes":[],"partners":["RAF1","AURKA","CIP2A","QPCT","BTRC","CD81"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"P01112","full_name":"GTPase HRas","aliases":["H-Ras-1","Ha-Ras","Transforming protein p21","c-H-ras","p21ras"],"length_aa":189,"mass_kda":21.3,"function":"Involved in the activation of Ras protein signal transduction (PubMed:22821884). Ras proteins bind GDP/GTP and possess intrinsic GTPase activity (PubMed:12740440, PubMed:14500341, PubMed:9020151)","subcellular_location":"Nucleus; Cytoplasm; Cytoplasm, perinuclear region","url":"https://www.uniprot.org/uniprotkb/P01112/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/HRAS","classification":"Not Classified","n_dependent_lines":40,"n_total_lines":1208,"dependency_fraction":0.033112582781456956},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/HRAS","total_profiled":1310},"omim":[{"mim_id":"621519","title":"SCAFFOLDING CK1-ANCHORING PROTEIN B; SACK1B","url":"https://www.omim.org/entry/621519"},{"mim_id":"621092","title":"IQ MOTIF-CONTAINING GTPase-ACTIVATING PROTEIN 3; IQGAP3","url":"https://www.omim.org/entry/621092"},{"mim_id":"621048","title":"POLO-LIKE KINASE 5, INACTIVE; PLK5","url":"https://www.omim.org/entry/621048"},{"mim_id":"620929","title":"MOB KINASE ACTIVATOR 3A; MOB3A","url":"https://www.omim.org/entry/620929"},{"mim_id":"620302","title":"WD REPEAT-CONTAINING PROTEIN 76; WDR76","url":"https://www.omim.org/entry/620302"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Cytosol","reliability":"Supported"},{"location":"Nucleoplasm","reliability":"Additional"},{"location":"Basal body","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"brain","ntpm":123.5}],"url":"https://www.proteinatlas.org/search/HRAS"},"hgnc":{"alias_symbol":[],"prev_symbol":["HRAS1"]},"alphafold":{"accession":"P01112","domains":[{"cath_id":"3.40.50.300","chopping":"1-170","consensus_level":"high","plddt":96.2983,"start":1,"end":170}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P01112","model_url":"https://alphafold.ebi.ac.uk/files/AF-P01112-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P01112-F1-predicted_aligned_error_v6.png","plddt_mean":91.94},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=HRAS","jax_strain_url":"https://www.jax.org/strain/search?query=HRAS"},"sequence":{"accession":"P01112","fasta_url":"https://rest.uniprot.org/uniprotkb/P01112.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P01112/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P01112"}},"corpus_meta":[{"pmid":"10980135","id":"PMC_10980135","title":"Mutations and copy number increase of HRAS in Spitz nevi with distinctive histopathological features.","date":"2000","source":"The American journal of pathology","url":"https://pubmed.ncbi.nlm.nih.gov/10980135","citation_count":325,"is_preprint":false},{"pmid":"7901201","id":"PMC_7901201","title":"Purification and properties of a palmitoyl-protein thioesterase that cleaves palmitate from H-Ras.","date":"1993","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/7901201","citation_count":295,"is_preprint":false},{"pmid":"22683711","id":"PMC_22683711","title":"Postzygotic HRAS and KRAS mutations cause nevus sebaceous and Schimmelpenning syndrome.","date":"2012","source":"Nature genetics","url":"https://pubmed.ncbi.nlm.nih.gov/22683711","citation_count":252,"is_preprint":false},{"pmid":"7541721","id":"PMC_7541721","title":"Overexpression of the hyaluronan receptor RHAMM is transforming and is also required for H-ras transformation.","date":"1995","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/7541721","citation_count":252,"is_preprint":false},{"pmid":"21072204","id":"PMC_21072204","title":"FGFR3, HRAS, KRAS, NRAS and PIK3CA mutations in bladder cancer and their potential as biomarkers for surveillance and therapy.","date":"2010","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/21072204","citation_count":251,"is_preprint":false},{"pmid":"3600660","id":"PMC_3600660","title":"The oncogenic forms of N-ras or H-ras prevent skeletal myoblast differentiation.","date":"1987","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/3600660","citation_count":218,"is_preprint":false},{"pmid":"19855393","id":"PMC_19855393","title":"Activating mutations in FGFR3 and HRAS reveal a shared genetic origin for congenital disorders and testicular tumors.","date":"2009","source":"Nature genetics","url":"https://pubmed.ncbi.nlm.nih.gov/19855393","citation_count":167,"is_preprint":false},{"pmid":"33750196","id":"PMC_33750196","title":"Tipifarnib in Head and Neck Squamous Cell Carcinoma With HRAS Mutations.","date":"2021","source":"Journal of clinical oncology : official journal of the American Society of Clinical Oncology","url":"https://pubmed.ncbi.nlm.nih.gov/33750196","citation_count":163,"is_preprint":false},{"pmid":"16329078","id":"PMC_16329078","title":"HRAS mutation analysis in Costello syndrome: genotype and phenotype correlation.","date":"2006","source":"American journal of medical genetics. Part A","url":"https://pubmed.ncbi.nlm.nih.gov/16329078","citation_count":146,"is_preprint":false},{"pmid":"24516166","id":"PMC_24516166","title":"H-Ras forms dimers on membrane surfaces via a protein-protein interface.","date":"2014","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/24516166","citation_count":133,"is_preprint":false},{"pmid":"23498955","id":"PMC_23498955","title":"HRas signal transduction promotes hepatitis C virus cell entry by triggering assembly of the host tetraspanin receptor complex.","date":"2013","source":"Cell host & microbe","url":"https://pubmed.ncbi.nlm.nih.gov/23498955","citation_count":133,"is_preprint":false},{"pmid":"15517309","id":"PMC_15517309","title":"Mutation analysis of the Ras pathway genes NRAS, HRAS, KRAS and BRAF in glioblastomas.","date":"2004","source":"Acta neuropathologica","url":"https://pubmed.ncbi.nlm.nih.gov/15517309","citation_count":129,"is_preprint":false},{"pmid":"15077182","id":"PMC_15077182","title":"Krüppel-like factor 5 mediates the transforming activity of oncogenic H-Ras.","date":"2004","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/15077182","citation_count":120,"is_preprint":false},{"pmid":"12077341","id":"PMC_12077341","title":"H-Ras signaling and K-Ras signaling are differentially dependent on endocytosis.","date":"2002","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/12077341","citation_count":119,"is_preprint":false},{"pmid":"29739933","id":"PMC_29739933","title":"Recurrent hotspot mutations in HRAS Q61 and PI3K-AKT pathway genes as drivers of breast adenomyoepitheliomas.","date":"2018","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/29739933","citation_count":114,"is_preprint":false},{"pmid":"19416908","id":"PMC_19416908","title":"Endogenous expression of Hras(G12V) induces developmental defects and neoplasms with copy number imbalances of the oncogene.","date":"2009","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/19416908","citation_count":113,"is_preprint":false},{"pmid":"18094044","id":"PMC_18094044","title":"A unique platform for H-Ras signaling involving clathrin-independent endocytosis.","date":"2007","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/18094044","citation_count":112,"is_preprint":false},{"pmid":"2247480","id":"PMC_2247480","title":"Overexpression of normal and mutated forms of HRAS induces orthotopic bladder invasion in a human transitional cell carcinoma.","date":"1990","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/2247480","citation_count":111,"is_preprint":false},{"pmid":"21170325","id":"PMC_21170325","title":"Kita driven expression of oncogenic HRAS leads to early onset and highly penetrant melanoma in zebrafish.","date":"2010","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/21170325","citation_count":111,"is_preprint":false},{"pmid":"2196280","id":"PMC_2196280","title":"H-ras protooncogene mutations in human thyroid neoplasms.","date":"1990","source":"The Journal of clinical endocrinology and metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/2196280","citation_count":108,"is_preprint":false},{"pmid":"22096025","id":"PMC_22096025","title":"Skin tumors induced by sorafenib; paradoxic RAS-RAF pathway activation and oncogenic mutations of HRAS, TP53, and TGFBR1.","date":"2011","source":"Clinical cancer research : an official journal of the American Association for Cancer Research","url":"https://pubmed.ncbi.nlm.nih.gov/22096025","citation_count":105,"is_preprint":false},{"pmid":"8290259","id":"PMC_8290259","title":"Subtraction cloning of H-rev107, a gene specifically expressed in H-ras resistant fibroblasts.","date":"1994","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/8290259","citation_count":105,"is_preprint":false},{"pmid":"2667764","id":"PMC_2667764","title":"Expression of the H-ras proto-oncogene is controlled by alternative splicing.","date":"1989","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/2667764","citation_count":104,"is_preprint":false},{"pmid":"31428295","id":"PMC_31428295","title":"Correlate the TP53 Mutation and the HRAS Mutation with Immune Signatures in Head and Neck Squamous Cell Cancer.","date":"2019","source":"Computational and structural biotechnology journal","url":"https://pubmed.ncbi.nlm.nih.gov/31428295","citation_count":96,"is_preprint":false},{"pmid":"24006476","id":"PMC_24006476","title":"Multilineage somatic activating mutations in HRAS and NRAS cause mosaic cutaneous and skeletal lesions, elevated FGF23 and hypophosphatemia.","date":"2013","source":"Human molecular genetics","url":"https://pubmed.ncbi.nlm.nih.gov/24006476","citation_count":95,"is_preprint":false},{"pmid":"21931711","id":"PMC_21931711","title":"G4-DNA formation in the HRAS promoter and rational design of decoy oligonucleotides for cancer therapy.","date":"2011","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/21931711","citation_count":92,"is_preprint":false},{"pmid":"23337891","id":"PMC_23337891","title":"Phacomatosis pigmentokeratotica is caused by a postzygotic HRAS mutation in a multipotent progenitor cell.","date":"2013","source":"The Journal of investigative dermatology","url":"https://pubmed.ncbi.nlm.nih.gov/23337891","citation_count":89,"is_preprint":false},{"pmid":"9799520","id":"PMC_9799520","title":"H-Ras peptide and protein substrates bind protein farnesyltransferase as an ionized thiolate.","date":"1998","source":"Biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/9799520","citation_count":89,"is_preprint":false},{"pmid":"19240121","id":"PMC_19240121","title":"H-Ras is degraded by Wnt/beta-catenin signaling via beta-TrCP-mediated polyubiquitylation.","date":"2009","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/19240121","citation_count":86,"is_preprint":false},{"pmid":"17250658","id":"PMC_17250658","title":"HRAS and the Costello syndrome.","date":"2007","source":"Clinical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/17250658","citation_count":83,"is_preprint":false},{"pmid":"20871217","id":"PMC_20871217","title":"HRAS-mutated Spitz tumors: A subtype of Spitz tumors with distinct features.","date":"2010","source":"The American journal of surgical pathology","url":"https://pubmed.ncbi.nlm.nih.gov/20871217","citation_count":82,"is_preprint":false},{"pmid":"27437771","id":"PMC_27437771","title":"Barriers to horizontal cell transformation by extracellular vesicles containing oncogenic H-ras.","date":"2016","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/27437771","citation_count":81,"is_preprint":false},{"pmid":"30994537","id":"PMC_30994537","title":"Diagnostic Significance of HRAS Mutations in Epithelial-Myoepithelial Carcinomas Exhibiting a Broad Histopathologic Spectrum.","date":"2019","source":"The American journal of surgical pathology","url":"https://pubmed.ncbi.nlm.nih.gov/30994537","citation_count":76,"is_preprint":false},{"pmid":"18268007","id":"PMC_18268007","title":"Activated Kras, but not Hras or Nras, may initiate tumors of endodermal origin via stem cell expansion.","date":"2008","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/18268007","citation_count":74,"is_preprint":false},{"pmid":"30787043","id":"PMC_30787043","title":"HRAS-driven cancer cells are vulnerable to TRPML1 inhibition.","date":"2019","source":"EMBO reports","url":"https://pubmed.ncbi.nlm.nih.gov/30787043","citation_count":73,"is_preprint":false},{"pmid":"7912510","id":"PMC_7912510","title":"H-ras oncogene mutation and human papillomavirus infection in oral carcinomas.","date":"1994","source":"Archives of otolaryngology--head & neck surgery","url":"https://pubmed.ncbi.nlm.nih.gov/7912510","citation_count":71,"is_preprint":false},{"pmid":"29760048","id":"PMC_29760048","title":"Tipifarnib Inhibits HRAS-Driven Dedifferentiated Thyroid Cancers.","date":"2018","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/29760048","citation_count":70,"is_preprint":false},{"pmid":"16969868","id":"PMC_16969868","title":"Somatic mosaicism for an HRAS mutation causes Costello syndrome.","date":"2006","source":"American journal of medical genetics. Part A","url":"https://pubmed.ncbi.nlm.nih.gov/16969868","citation_count":64,"is_preprint":false},{"pmid":"19438459","id":"PMC_19438459","title":"BRAF, NRAS and HRAS mutations in spitzoid tumours and their possible pathogenetic significance.","date":"2009","source":"The British journal of dermatology","url":"https://pubmed.ncbi.nlm.nih.gov/19438459","citation_count":63,"is_preprint":false},{"pmid":"24277618","id":"PMC_24277618","title":"HRAS mutations in epithelial-myoepithelial carcinoma.","date":"2013","source":"Head and neck pathology","url":"https://pubmed.ncbi.nlm.nih.gov/24277618","citation_count":61,"is_preprint":false},{"pmid":"11007040","id":"PMC_11007040","title":"Molecular abnormalities of p53, MDM2, and H-ras in synovial sarcoma.","date":"2000","source":"Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc","url":"https://pubmed.ncbi.nlm.nih.gov/11007040","citation_count":60,"is_preprint":false},{"pmid":"24341335","id":"PMC_24341335","title":"PIK3CA, HRAS and PTEN in human papillomavirus positive oropharyngeal squamous cell carcinoma.","date":"2013","source":"BMC cancer","url":"https://pubmed.ncbi.nlm.nih.gov/24341335","citation_count":56,"is_preprint":false},{"pmid":"16568090","id":"PMC_16568090","title":"Oncogenic HRAS suppresses clusterin expression through promoter hypermethylation.","date":"2006","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/16568090","citation_count":56,"is_preprint":false},{"pmid":"25072932","id":"PMC_25072932","title":"CDKN2A(p16) and HRAS are frequently mutated in vulvar squamous cell carcinoma.","date":"2014","source":"Gynecologic oncology","url":"https://pubmed.ncbi.nlm.nih.gov/25072932","citation_count":53,"is_preprint":false},{"pmid":"23868059","id":"PMC_23868059","title":"H-Ras transfers from B to T cells via tunneling nanotubes.","date":"2013","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/23868059","citation_count":53,"is_preprint":false},{"pmid":"26674223","id":"PMC_26674223","title":"GC-elements controlling HRAS transcription form i-motif structures unfolded by heterogeneous ribonucleoprotein particle A1.","date":"2015","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/26674223","citation_count":51,"is_preprint":false},{"pmid":"12730683","id":"PMC_12730683","title":"Identification of H-Ras, RhoA, Rac1 and Cdc42 responsive genes.","date":"2003","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/12730683","citation_count":51,"is_preprint":false},{"pmid":"23512660","id":"PMC_23512660","title":"Inflammation and Hras signaling control epithelial-mesenchymal transition during skin tumor progression.","date":"2013","source":"Genes & development","url":"https://pubmed.ncbi.nlm.nih.gov/23512660","citation_count":50,"is_preprint":false},{"pmid":"9324018","id":"PMC_9324018","title":"H-ras oncogene point mutations in arthritic synovium.","date":"1997","source":"Arthritis and rheumatism","url":"https://pubmed.ncbi.nlm.nih.gov/9324018","citation_count":50,"is_preprint":false},{"pmid":"22085642","id":"PMC_22085642","title":"Oxidation of HRas cysteine thiols by metabolic stress prevents palmitoylation in vivo and contributes to endothelial cell apoptosis.","date":"2011","source":"FASEB journal : official publication of the Federation of American Societies for Experimental Biology","url":"https://pubmed.ncbi.nlm.nih.gov/22085642","citation_count":50,"is_preprint":false},{"pmid":"26858456","id":"PMC_26858456","title":"Endocytosis separates EGF receptors from endogenous fluorescently labeled HRas and diminishes receptor signaling to MAP kinases in endosomes.","date":"2016","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/26858456","citation_count":47,"is_preprint":false},{"pmid":"31534544","id":"PMC_31534544","title":"DNA methylation-regulated QPCT promotes sunitinib resistance by increasing HRAS stability in renal cell carcinoma.","date":"2019","source":"Theranostics","url":"https://pubmed.ncbi.nlm.nih.gov/31534544","citation_count":46,"is_preprint":false},{"pmid":"26544513","id":"PMC_26544513","title":"Mutant HRAS as novel target for MEK and mTOR inhibitors.","date":"2015","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/26544513","citation_count":45,"is_preprint":false},{"pmid":"25458953","id":"PMC_25458953","title":"CIP2A cooperates with H-Ras to promote epithelial-mesenchymal transition in cervical-cancer progression.","date":"2014","source":"Cancer letters","url":"https://pubmed.ncbi.nlm.nih.gov/25458953","citation_count":45,"is_preprint":false},{"pmid":"27067779","id":"PMC_27067779","title":"Porocarcinomas harbor recurrent HRAS-activating mutations and tumor suppressor inactivating mutations.","date":"2016","source":"Human pathology","url":"https://pubmed.ncbi.nlm.nih.gov/27067779","citation_count":45,"is_preprint":false},{"pmid":"32557577","id":"PMC_32557577","title":"Tipifarnib in recurrent, metastatic HRAS-mutant salivary gland cancer.","date":"2020","source":"Cancer","url":"https://pubmed.ncbi.nlm.nih.gov/32557577","citation_count":43,"is_preprint":false},{"pmid":"22199277","id":"PMC_22199277","title":"Mutational screening of RET, HRAS, KRAS, NRAS, BRAF, AKT1, and CTNNB1 in medullary thyroid carcinoma.","date":"2011","source":"Anticancer research","url":"https://pubmed.ncbi.nlm.nih.gov/22199277","citation_count":42,"is_preprint":false},{"pmid":"27725900","id":"PMC_27725900","title":"Identification of HRAS as cancer-promoting gene in gastric carcinoma cell aggressiveness.","date":"2016","source":"American journal of cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/27725900","citation_count":41,"is_preprint":false},{"pmid":"17979197","id":"PMC_17979197","title":"Mutation analysis in Costello syndrome: functional and structural characterization of the HRAS p.Lys117Arg mutation.","date":"2008","source":"Human mutation","url":"https://pubmed.ncbi.nlm.nih.gov/17979197","citation_count":40,"is_preprint":false},{"pmid":"35459782","id":"PMC_35459782","title":"Targeting farnesylation as a novel therapeutic approach in HRAS-mutant rhabdomyosarcoma.","date":"2022","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/35459782","citation_count":39,"is_preprint":false},{"pmid":"38452464","id":"PMC_38452464","title":"Polystyrene microplastics induce anxiety via HRAS derived PERK-NF-κB pathway.","date":"2024","source":"Environment international","url":"https://pubmed.ncbi.nlm.nih.gov/38452464","citation_count":39,"is_preprint":false},{"pmid":"29901113","id":"PMC_29901113","title":"HRAS as a potential therapeutic target of salirasib RAS inhibitor in bladder cancer.","date":"2018","source":"International journal of oncology","url":"https://pubmed.ncbi.nlm.nih.gov/29901113","citation_count":39,"is_preprint":false},{"pmid":"18355852","id":"PMC_18355852","title":"PIK3CA, HRAS and KRAS gene mutations in human penile cancer.","date":"2008","source":"The Journal of urology","url":"https://pubmed.ncbi.nlm.nih.gov/18355852","citation_count":38,"is_preprint":false},{"pmid":"30865897","id":"PMC_30865897","title":"An Integrated Global Analysis of Compartmentalized HRAS Signaling.","date":"2019","source":"Cell reports","url":"https://pubmed.ncbi.nlm.nih.gov/30865897","citation_count":37,"is_preprint":false},{"pmid":"18030338","id":"PMC_18030338","title":"Intercellular transfer of oncogenic H-Ras at the immunological synapse.","date":"2007","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/18030338","citation_count":37,"is_preprint":false},{"pmid":"19371735","id":"PMC_19371735","title":"Costello syndrome H-Ras alleles regulate cortical development.","date":"2009","source":"Developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/19371735","citation_count":36,"is_preprint":false},{"pmid":"25097040","id":"PMC_25097040","title":"HRAS mutations are frequent in inverted urothelial neoplasms.","date":"2014","source":"Human pathology","url":"https://pubmed.ncbi.nlm.nih.gov/25097040","citation_count":34,"is_preprint":false},{"pmid":"23599145","id":"PMC_23599145","title":"Identification of HRAS mutations and absence of GNAQ or GNA11 mutations in deep penetrating nevi.","date":"2013","source":"Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc","url":"https://pubmed.ncbi.nlm.nih.gov/23599145","citation_count":32,"is_preprint":false},{"pmid":"15087391","id":"PMC_15087391","title":"K-Ras and H-Ras activation promote distinct consequences on endometrial cell survival.","date":"2004","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/15087391","citation_count":32,"is_preprint":false},{"pmid":"31887226","id":"PMC_31887226","title":"Immunohistochemical assessment of HRAS Q61R mutations in breast adenomyoepitheliomas.","date":"2020","source":"Histopathology","url":"https://pubmed.ncbi.nlm.nih.gov/31887226","citation_count":30,"is_preprint":false},{"pmid":"26872011","id":"PMC_26872011","title":"Frequent HRAS Mutations in Malignant Ectomesenchymoma: Overlapping Genetic Abnormalities With Embryonal Rhabdomyosarcoma.","date":"2016","source":"The American journal of surgical pathology","url":"https://pubmed.ncbi.nlm.nih.gov/26872011","citation_count":30,"is_preprint":false},{"pmid":"37339176","id":"PMC_37339176","title":"Tipifarnib Potentiates the Antitumor Effects of PI3Kα Inhibition in PIK3CA- and HRAS-Dysregulated HNSCC via Convergent Inhibition of mTOR Activity.","date":"2023","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/37339176","citation_count":30,"is_preprint":false},{"pmid":"31637524","id":"PMC_31637524","title":"Arteriovenous malformation associated with a HRAS mutation.","date":"2019","source":"Human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/31637524","citation_count":30,"is_preprint":false},{"pmid":"30115483","id":"PMC_30115483","title":"ERK-TSC2 signalling in constitutively-active HRAS mutant HNSCC cells promotes resistance to PI3K inhibition.","date":"2018","source":"Oral oncology","url":"https://pubmed.ncbi.nlm.nih.gov/30115483","citation_count":30,"is_preprint":false},{"pmid":"26773571","id":"PMC_26773571","title":"HRAS mutation prevalence and associated expression patterns in pheochromocytoma.","date":"2016","source":"Genes, chromosomes & cancer","url":"https://pubmed.ncbi.nlm.nih.gov/26773571","citation_count":29,"is_preprint":false},{"pmid":"24169525","id":"PMC_24169525","title":"HRAS mutations in bladder cancer at an early age and the possible association with the Costello Syndrome.","date":"2013","source":"European journal of human genetics : EJHG","url":"https://pubmed.ncbi.nlm.nih.gov/24169525","citation_count":28,"is_preprint":false},{"pmid":"36603172","id":"PMC_36603172","title":"HRAS Mutations Define a Distinct Subgroup in Head and Neck Squamous Cell Carcinoma.","date":"2023","source":"JCO precision oncology","url":"https://pubmed.ncbi.nlm.nih.gov/36603172","citation_count":27,"is_preprint":false},{"pmid":"27175596","id":"PMC_27175596","title":"MAPK activation and HRAS mutation identified in pituitary spindle cell oncocytoma.","date":"2016","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/27175596","citation_count":26,"is_preprint":false},{"pmid":"32970285","id":"PMC_32970285","title":"CircZNF609 promotes cell proliferation, migration, invasion, and glycolysis in nasopharyngeal carcinoma through regulating HRAS via miR-338-3p.","date":"2020","source":"Molecular and cellular biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/32970285","citation_count":26,"is_preprint":false},{"pmid":"27238285","id":"PMC_27238285","title":"H-ras Inhibits the Hippo Pathway by Promoting Mst1/Mst2 Heterodimerization.","date":"2016","source":"Current biology : CB","url":"https://pubmed.ncbi.nlm.nih.gov/27238285","citation_count":26,"is_preprint":false},{"pmid":"31960612","id":"PMC_31960612","title":"Frequent KRAS and HRAS mutations in squamous cell papillomas of the head and neck.","date":"2020","source":"The journal of pathology. Clinical research","url":"https://pubmed.ncbi.nlm.nih.gov/31960612","citation_count":25,"is_preprint":false},{"pmid":"22820081","id":"PMC_22820081","title":"The mutational spectrum of HRAS, KRAS, NRAS and FGFR3 genes in bladder cancer.","date":"2011","source":"Cancer biomarkers : section A of Disease markers","url":"https://pubmed.ncbi.nlm.nih.gov/22820081","citation_count":25,"is_preprint":false},{"pmid":"36753744","id":"PMC_36753744","title":"Mutated HRAS Activates YAP1-AXL Signaling to Drive Metastasis of Head and Neck Cancer.","date":"2023","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/36753744","citation_count":23,"is_preprint":false},{"pmid":"2406716","id":"PMC_2406716","title":"Nontumorigenic squamous cell carcinoma line converted to tumorigenicity with methyl methanesulfonate without activation of HRAS or MYC.","date":"1990","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/2406716","citation_count":22,"is_preprint":false},{"pmid":"27503857","id":"PMC_27503857","title":"SPRED1 Interferes with K-ras but Not H-ras Membrane Anchorage and Signaling.","date":"2016","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/27503857","citation_count":22,"is_preprint":false},{"pmid":"28177880","id":"PMC_28177880","title":"Aurora kinase A interacts with H-Ras and potentiates Ras-MAPK signaling.","date":"2017","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/28177880","citation_count":21,"is_preprint":false},{"pmid":"30191474","id":"PMC_30191474","title":"The efficacy of HRAS and CDK4/6 inhibitors in anaplastic thyroid cancer cell lines.","date":"2018","source":"Journal of endocrinological investigation","url":"https://pubmed.ncbi.nlm.nih.gov/30191474","citation_count":20,"is_preprint":false},{"pmid":"20884686","id":"PMC_20884686","title":"The TGF-beta co-receptor endoglin modulates the expression and transforming potential of H-Ras.","date":"2010","source":"Carcinogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/20884686","citation_count":20,"is_preprint":false},{"pmid":"35230976","id":"PMC_35230976","title":"HRAS germline mutations impair LKB1/AMPK signaling and mitochondrial homeostasis in Costello syndrome models.","date":"2022","source":"The Journal of clinical investigation","url":"https://pubmed.ncbi.nlm.nih.gov/35230976","citation_count":19,"is_preprint":false},{"pmid":"21868531","id":"PMC_21868531","title":"H-ras up-regulates expression of BNIP3.","date":"2011","source":"Anticancer research","url":"https://pubmed.ncbi.nlm.nih.gov/21868531","citation_count":19,"is_preprint":false},{"pmid":"8690291","id":"PMC_8690291","title":"Expression and mutation of H-ras in uterine cervical cancer.","date":"1996","source":"Gynecologic oncology","url":"https://pubmed.ncbi.nlm.nih.gov/8690291","citation_count":19,"is_preprint":false},{"pmid":"25026275","id":"PMC_25026275","title":"H-Ras regulation of TRAIL death receptor mediated apoptosis.","date":"2014","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/25026275","citation_count":19,"is_preprint":false},{"pmid":"27798864","id":"PMC_27798864","title":"Inhibition of Galectin-1 Sensitizes HRAS-driven Tumor Growth to Rapamycin Treatment.","date":"2016","source":"Anticancer research","url":"https://pubmed.ncbi.nlm.nih.gov/27798864","citation_count":19,"is_preprint":false},{"pmid":"1535685","id":"PMC_1535685","title":"Novel revertants of H-ras oncogene-transformed R6-PKC3 cells.","date":"1992","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/1535685","citation_count":19,"is_preprint":false},{"pmid":"36996116","id":"PMC_36996116","title":"Epitranscriptic regulation of HRAS by N6-methyladenosine drives tumor progression.","date":"2023","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/36996116","citation_count":18,"is_preprint":false},{"pmid":"29097261","id":"PMC_29097261","title":"Calmodulin and IQGAP1 activation of PI3Kα and Akt in KRAS, HRAS and NRAS-driven cancers.","date":"2017","source":"Biochimica et biophysica acta. Molecular basis of disease","url":"https://pubmed.ncbi.nlm.nih.gov/29097261","citation_count":16,"is_preprint":false},{"pmid":"19191262","id":"PMC_19191262","title":"Increased genomic instability and altered chromosomal protein phosphorylation timing in HRAS-transformed mouse fibroblasts.","date":"2009","source":"Genes, chromosomes & cancer","url":"https://pubmed.ncbi.nlm.nih.gov/19191262","citation_count":16,"is_preprint":false},{"pmid":"34496925","id":"PMC_34496925","title":"HRAS is a therapeutic target in malignant chemo-resistant adenomyoepithelioma of the breast.","date":"2021","source":"Journal of hematology & oncology","url":"https://pubmed.ncbi.nlm.nih.gov/34496925","citation_count":16,"is_preprint":false},{"pmid":"36995904","id":"PMC_36995904","title":"Crystal Structure of an i-Motif from the HRAS Oncogene Promoter.","date":"2023","source":"Angewandte Chemie (International ed. in English)","url":"https://pubmed.ncbi.nlm.nih.gov/36995904","citation_count":16,"is_preprint":false},{"pmid":"34553752","id":"PMC_34553752","title":"MEK-inhibitor-mediated rescue of skeletal myopathy caused by activating Hras mutation in a Costello syndrome mouse model.","date":"2021","source":"Disease models & mechanisms","url":"https://pubmed.ncbi.nlm.nih.gov/34553752","citation_count":16,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":52120,"output_tokens":9476,"usd":0.14925,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":19630,"output_tokens":7320,"usd":0.140575,"stage2_stop_reason":"end_turn"},"total_usd":0.289825,"stage1_batch_id":"msgbatch_01DUPACZi7AAWwuZiUjseQF4","stage2_batch_id":"msgbatch_018yt7chE3f48PEFtDwtj6AE","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1993,\n      \"finding\": \"A novel 37-kDa palmitoyl-protein thioesterase purified from bovine brain cytosol enzymatically removes palmitate from H-Ras. The thioesterase recognizes H-Ras only in its native conformation (bound to Mg2+ and guanine nucleotide), and it is sensitive to diethyl pyrocarbonate but insensitive to PMSF and NEM. Palmitoylated alpha subunits of heterotrimeric G proteins are also substrates.\",\n      \"method\": \"Enzyme purification to homogeneity, in vitro thioesterase assay with [3H]palmitate-labeled H-Ras produced in baculovirus, chemical inhibitor profiling\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — reconstituted in vitro enzymatic assay with purified enzyme and defined substrate, multiple inhibitor controls, single rigorous study\",\n      \"pmids\": [\"7901201\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1998,\n      \"finding\": \"H-Ras peptide and protein substrates bind protein farnesyltransferase (FTase) as an ionized thiolate: metal coordination of the substrate cysteine sulfur by the catalytic zinc ion lowers the pKa of the thiol (pKa ~6.3), generating a bound thiolate at physiological pH. The two substrates (H-Ras CAAX peptide and FPP) bind FTase synergistically, with affinity enhanced ~70-fold in the ternary complex.\",\n      \"method\": \"Fluorescence binding assays, pH-dependent affinity measurements, optical absorption spectroscopy of Co2+-substituted FTase, in vitro biochemical assay\",\n      \"journal\": \"Biochemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro reconstitution with purified enzyme, multiple orthogonal spectroscopic methods, mechanistic detail on catalytic zinc coordination\",\n      \"pmids\": [\"9799520\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1989,\n      \"finding\": \"Alternative splicing of the H-ras pre-mRNA normally suppresses p21 H-Ras expression: a negative-acting element in the last intron functions as an alternative exon; transcripts containing this exon are produced at low abundance (due to instability/defective processing) and are predicted to lack transforming potential. Mutations that prevent inclusion of this alternative exon increase H-ras expression and transforming efficiency.\",\n      \"method\": \"Gene reconstruction experiments, S1 nuclease analysis, mutational analysis of intronic elements, transcript identification\",\n      \"journal\": \"Cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — multiple orthogonal molecular biology methods (gene reconstruction, S1 analysis, mutational scanning) in a single rigorous study establishing the splicing control mechanism\",\n      \"pmids\": [\"2667764\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1995,\n      \"finding\": \"RHAMM (hyaluronan receptor) acts downstream of H-Ras in the transformation pathway: fibroblasts with a dominant suppressor mutant of RHAMM revert H-ras-transformed fibrosarcomas to a nontumorigenic, nonmetastatic phenotype, and antisense suppression of RHAMM renders fibroblasts resistant to ras transformation. Loss of functional RHAMM ablates focal adhesion kinase phosphorylation signaling within focal adhesions, preventing turnover in response to hyaluronan.\",\n      \"method\": \"Dominant suppressor transfection, antisense transfection, soft agar assays, in vivo tumorigenicity and metastasis assays, focal adhesion kinase phosphorylation analysis, genetic epistasis\",\n      \"journal\": \"Cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic epistasis (dominant suppressor + antisense) combined with defined biochemical readout (FAK phosphorylation) and in vivo tumor assays\",\n      \"pmids\": [\"7541721\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"H-Ras signaling through the Raf/MEK/MAPK cascade requires endocytosis and endocytic recycling, whereas K-Ras signaling does not. Dominant-interfering dynamin-K44A selectively blocked H-Ras- but not K-Ras-mediated Raf-1 activation and PC12 differentiation. Wild-type Rab5 (stimulating endocytosis and recycling) potentiated H-Ras/Raf-1 signaling, while Rab5-Q79L (endocytosis without recycling) redistributed active H-Ras to enlarged endosomes and inhibited H-Ras/Raf-1 activation. H-Ras (but not K-Ras) signaling also required PI3K activity.\",\n      \"method\": \"Dominant-interfering dynamin expression, Rab5 mutant overexpression, Raf-1 kinase activity assay, subcellular fractionation/localization, PC12 differentiation assay\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple genetic tools (dominant-negative dynamin, activating and non-recycling Rab5 mutants) with isoform-specific readouts in two cell systems\",\n      \"pmids\": [\"12077341\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"H-Ras enters cells via clathrin-independent endocytosis (CIE): it co-localizes with MHC class I CIE cargo and is sequestered in Arf6-Q67L vacuoles. Activated H-RasG12V induces macropinocytosis; incoming macropinosomes sequentially contain PIP2/PIP3, then PIP3/Rab5, then Rab5 alone. Arf6-Q67L traps H-Ras signaling in the PIP2/PIP3 stage, recruiting active ERK and Akt but not Rab5.\",\n      \"method\": \"Live fluorescence imaging, PH-domain reporters for phosphoinositides, Arf6 and Rab5 mutant expression, co-localization with CIE cargo markers\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — live imaging with lipid reporters and genetic tools in a single lab; clear mechanistic staging of endosomal H-Ras signaling\",\n      \"pmids\": [\"18094044\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"H-Ras protein is degraded via beta-TrCP-mediated polyubiquitylation and proteasomal degradation. H-Ras interacts with the WD40 domain of beta-TrCP; this ubiquitylation is stimulated by Axin or APC and inhibited by Wnt3a. In vivo, intravenous Wnt3a injection in mice reduces Ras levels in intestinal tissue, establishing Wnt/beta-catenin signaling as a regulator of H-Ras stability and providing a mechanistic basis for crosstalk between Wnt and Ras-ERK pathways.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitylation assay, proteasome inhibitor experiments, Axin/APC/Wnt3a gain- and loss-of-function, in vivo Wnt3a injection in mice\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP mapping to WD40 domain, ubiquitylation assay, multiple gain/loss-of-function reagents, in vivo validation\",\n      \"pmids\": [\"19240121\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Metabolic stress (high palmitate/glucose) oxidizes H-Ras cysteine thiols at Cys181/184, detected by MALDI-TOF/TOF mass spectrometry, which prevents palmitoylation. Loss of palmitoylation shifts H-Ras from plasma membrane to Golgi, decreases growth factor-stimulated ERK phosphorylation (~84% reduction), and increases apoptotic signaling; these effects are prevented by wild-type but not C181/184S H-Ras, establishing oxidation of palmitoylation-site cysteines as the causal mechanism.\",\n      \"method\": \"MALDI-TOF/TOF mass spectrometry of Cys181/184, palmitoylation assay, subcellular fractionation, ERK phosphorylation measurement, apoptosis assay, WT vs. C181/184S mutant rescue, MnSOD overexpression\",\n      \"journal\": \"FASEB journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — direct mass spectrometric identification of oxidized cysteines, site-specific mutagenesis (C181/184S), and multiple orthogonal functional readouts in a single study\",\n      \"pmids\": [\"22085642\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"H-Ras forms dimers on membrane surfaces via a protein-protein interface involving the switch II region (near the nucleotide binding cleft). A Y64A point mutation in switch II abolishes dimer formation. Dimerization requires the membrane surface and does not occur in solution at comparable densities; it does not require lipid anchor clustering. The 2D dimerization Kd is ~1 × 10^3 molecules/μm^2, and no higher-order oligomers are observed.\",\n      \"method\": \"Fluorescence correlation spectroscopy, photon counting histogram analysis, time-resolved fluorescence anisotropy, single-molecule tracking, step photobleaching, supported lipid bilayer reconstitution, site-directed mutagenesis (Y64A)\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — multiple orthogonal single-molecule and ensemble biophysical methods on reconstituted membranes, mutagenesis validation, quantitative Kd measurement\",\n      \"pmids\": [\"24516166\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"HRas GTPase, activated downstream of EGFR signaling, is required for HCV entry into hepatocytes. Proteomic analysis showed HRas associates with tetraspanin CD81, claudin-1, integrin β1, and Rap2B in hepatocyte membranes. HRas signaling mediates lateral membrane diffusion of CD81, enabling tetraspanin receptor complex assembly necessary for HCV entry.\",\n      \"method\": \"Proteomic/MS analysis of HRas-associated complex, RNAi knockdown of HRas, lateral membrane diffusion assays, HCV infection assay, Co-IP\",\n      \"journal\": \"Cell host & microbe\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — MS-based interactome identification, RNAi functional validation, lateral diffusion measurement, and infection assay provide multiple orthogonal lines of evidence\",\n      \"pmids\": [\"23498955\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"In H-Ras-transformed cells, elevated MAPK activity (from H-Ras) transcriptionally induces Egr1, which in turn upregulates KLF5; KLF5 then activates cyclin D1 expression. MEK inhibitors reduce KLF5 and abolish anchorage-independent growth. siRNA knockdown of KLF5 in H-Ras-transformed cells reduces proliferation and colony formation, establishing KLF5 as a downstream mediator of H-Ras oncogenic transformation via the MEK/ERK → Egr1 → KLF5 → cyclin D1 axis.\",\n      \"method\": \"MEK inhibitor treatment, siRNA knockdown of KLF5 and Egr1, soft agar colony formation, proliferation assays, qRT-PCR, Western blot\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — RNAi epistasis at two nodes (Egr1, KLF5) with defined phenotypic readouts; single lab\",\n      \"pmids\": [\"15077182\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"H-ras inhibits the Hippo tumor suppressor pathway by promoting formation of inactive Mst1/Mst2 heterodimers via an ERK-dependent mechanism. Mst1/Mst2 heterodimers (mediated by SARAH domains) have much lower kinase activity than homodimers. Cells lacking Mst1 (unable to form heterodimers) are resistant to H-ras-mediated transformation and maintain active Hippo signaling.\",\n      \"method\": \"Co-immunoprecipitation of Mst1/Mst2 heterodimers, in vitro kinase assay comparing homo- vs. heterodimer activity, MEK inhibitor epistasis, Mst1 knockout cells, focus formation assay\",\n      \"journal\": \"Current biology : CB\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — Co-IP demonstrating heterodimer formation, in vitro kinase activity comparison, genetic KO epistasis, and transformation assay provide multiple orthogonal mechanistic validations\",\n      \"pmids\": [\"27238285\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Aurora kinase A (Aurora A) physically interacts with H-Ras through its kinase domain binding the N-terminal domain of H-Ras; Aurora A, H-Ras, and Raf-1 exist in a trimeric complex. Aurora A enhances H-Ras binding to Raf-1 and potentiates H-Ras-mediated MAPK signaling in an active H-Ras-dependent manner.\",\n      \"method\": \"Co-immunoprecipitation, domain-mapping pulldown assays, MAPK signaling readouts, overexpression of Aurora A with active/inactive H-Ras\",\n      \"journal\": \"Oncotarget\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal Co-IP with domain mapping and functional MAPK readout; single lab\",\n      \"pmids\": [\"28177880\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"H-Ras transfers between cells via tunneling nanotubes (TNTs) connecting B and T cells. GFP-H-Ras diffuses freely in the TNT membrane (FRAP), and PM patches enriched in GFP-H-Ras and CD86 transfer to T-cell surfaces. Transfer is dependent on normal post-translational lipidation (palmitoylation/farnesylation) of H-Ras for plasma membrane anchorage.\",\n      \"method\": \"Optical tweezers cell trapping, 4D spinning-disk confocal live imaging, FRAP on TNT membranes, FACS, lipidation-defective mutant H-Ras controls\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — live imaging and FRAP with lipidation-mutant controls; single lab, single study\",\n      \"pmids\": [\"23868059\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"The HRAS p.Lys117Arg mutation (Costello syndrome) constitutively activates the RAS/MAPK pathway through a mechanism distinct from codon 12/13 mutations: it increases the nucleotide dissociation rate ~80-fold while retaining normal intrinsic GTP hydrolysis and GAP responsiveness. The crystal structure of HRAS p.Lys117Arg shows an altered side-chain interaction pattern that unfavorably affects nucleotide binding.\",\n      \"method\": \"Fluorescence nucleotide exchange/dissociation assays, GTPase activity assay, GAP-stimulated hydrolysis assay, X-ray crystallography of recombinant mutant protein\",\n      \"journal\": \"Human mutation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structure plus multiple in vitro biochemical assays (exchange rate, hydrolysis, GAP responsiveness) on recombinant mutant protein\",\n      \"pmids\": [\"17979197\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"TRPML1 (encoded by MCOLN1) maintains oncogenic HRAS in signaling-competent nanoclusters at the plasma membrane by mediating cholesterol de-esterification and transport. TRPML1 inhibition disrupts cholesterol distribution, attenuates HRAS nanoclustering and plasma membrane abundance, and reduces ERK phosphorylation and proliferation selectively in HRAS-mutant cancer cells.\",\n      \"method\": \"MCOLN1 knockdown and pharmacological TRPML1 inhibition, cholesterol distribution assays, HRAS nanoclustering quantification, ERK phosphorylation, proliferation assays in HRAS-mutant vs. wild-type cell lines\",\n      \"journal\": \"EMBO reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KD and pharmacological inhibition with mechanistic cholesterol/nanoclustering readouts; single lab\",\n      \"pmids\": [\"30787043\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Endogenous H-Ras (mVenus-HRas knock-in) is primarily located at the plasma membrane and in small amounts in tubular recycling endosomes; EGF stimulation causes fast but transient HRas activation. EGFR endocytosis physically separates EGFR-Grb2 complexes from HRas, terminating Ras-mediated signaling, while sustained minimal ERK activation is maintained by a small pool of active EGFRs remaining at the plasma membrane.\",\n      \"method\": \"CRISPR/gene-editing to tag endogenous HRas with mVenus, live fluorescence imaging, subcellular colocalization, EGFR/Grb2 endosomal tracking, surface biotinylation, selective inactivation of surface EGFRs, ERK/MEK phosphorylation time courses\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — endogenous tagging via gene editing, multiple imaging and biochemical readouts, and specific surface EGFR inactivation experiments in a single rigorous study\",\n      \"pmids\": [\"26858456\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Compartment-specific HRAS signaling: HRAS signaling is strongest from the cell membrane (highest kinase activation) but regulates the largest number of genes from the endoplasmic reticulum. Signaling from the Golgi apparatus activates TP53-dependent cell survival. Different subcellular locations generate distinct protein interaction profiles and kinase activation patterns that differentially regulate gene transcription.\",\n      \"method\": \"Compartment-targeted HRAS constructs, phosphoproteomics, protein interaction proteomics, transcriptomics, integrated network analysis\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple omics methods with compartment-targeted constructs; single lab, complex integrated analysis\",\n      \"pmids\": [\"30865897\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"QPCT binds directly to HRAS and attenuates its ubiquitination, thereby increasing HRAS protein stability and activating the ERK pathway. This interaction was identified by human proteome microarray and validated by co-IP and confocal colocalization. QPCT upregulation (driven by reduced promoter methylation) confers sunitinib resistance in renal cell carcinoma via HRAS stabilization.\",\n      \"method\": \"Human proteome microarray, co-immunoprecipitation, confocal colocalization, ubiquitination assay, siRNA knockdown, overexpression, ERK pathway readouts\",\n      \"journal\": \"Theranostics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — proteome microarray binding identification confirmed by Co-IP and colocalization, with mechanistic ubiquitination assay; single lab\",\n      \"pmids\": [\"31534544\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"m6A modification at three specific sites in the HRAS 3' UTR (but not KRAS or NRAS 3' UTR) promotes H-Ras protein expression by enhancing translational elongation. This modification is regulated by the m6A eraser FTO (which removes the modification) and read by YTHDF1 (which promotes translation); YTHDF2 and YTHDF3 are not involved. Targeting HRAS m6A modification decreases cancer cell proliferation and metastasis.\",\n      \"method\": \"m6A sequencing, luciferase reporter assays with mutated m6A sites, YTHDF1/2/3 and FTO knockdown/overexpression, polysome profiling for translational elongation, proliferation and metastasis assays\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple writer/reader/eraser perturbation experiments with specific 3' UTR site mapping; single lab\",\n      \"pmids\": [\"36996116\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"The HRAS promoter contains two G-rich elements (hras-1 and hras-2) that fold into G-quadruplex structures acting as transcriptional repressors. MAZ binds both quadruplexes; Sp1 binds only qhras-1. hnRNP A1 binds the complementary C-rich i-motif structures and unfolds them (demonstrated by FRET and CD), and is required for HRAS transcriptional activation (knockdown reduces transcript to ~44%).\",\n      \"method\": \"Chromatin immunoprecipitation (ChIP), EMSA, promoter mutagenesis (blocking or stabilizing quadruplexes), transcription assays, quadruplex-stabilizing ligands, RNAi knockdown\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP in cells combined with in vitro EMSA and mutagenesis; multiple regulatory elements characterized; single lab\",\n      \"pmids\": [\"21931711\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"hnRNP A1 binds the C-rich i-motif structures in the HRAS promoter GC-elements (hras-1 and hras-2), unfolds them upon binding (demonstrated by FRET and CD), and is necessary for HRAS transcriptional activation. Knockdown of hnRNP A1 reduces HRAS transcript to 44% of control. Sequestration of i-motif-binding proteins by decoy oligonucleotides significantly inhibits HRAS transcription.\",\n      \"method\": \"ChIP, EMSA, FRET, CD spectroscopy, shRNA knockdown of hnRNP A1, decoy oligonucleotide competition\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — biophysical (FRET, CD) and cell-based (ChIP, shRNA) methods concordant; single lab replicating and extending prior findings\",\n      \"pmids\": [\"26674223\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"The crystal structure of the HRAS oncogene promoter i-motif (iHRAS) was solved at 1.77 Å resolution. iHRAS folds into a double hairpin forming an antiparallel i-motif dimer capped by loops; six C-C+ base pairs form each i-motif core, extended by a G-G base pair and cytosine stacking. Extensive canonical and non-canonical base pairing stabilizes the connecting region and loops.\",\n      \"method\": \"X-ray crystallography at 1.77 Å resolution\",\n      \"journal\": \"Angewandte Chemie (International ed. in English)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — atomic resolution crystal structure with full structural determination\",\n      \"pmids\": [\"36995904\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Endoglin (TGF-β co-receptor) inhibits H-Ras expression and its oncogenic transformation capacity. TGF-β1 increases H-Ras mRNA and protein via ALK5 and the Ras-MAPK pathway (Smad4-independent); endoglin attenuates this TGF-β1-mediated induction. Endoglin also inhibits Ras-GTP levels, phospho-MEK, and phospho-ERK in H-Ras oncogene-expressing cells, and suppresses transformation by H-RasQ61K and H-RasG12V in NIH3T3 focus formation assays.\",\n      \"method\": \"H-Ras promoter transactivation assay, Northern/Western blot, Ras-GTP pulldown, MEK/ERK phosphorylation, NIH3T3 focus formation assay, ALK5 inhibitor epistasis\",\n      \"journal\": \"Carcinogenesis\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple biochemical readouts and functional transformation assay; single lab\",\n      \"pmids\": [\"20884686\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"CIP2A physically associates with H-Ras, leading to activation of the MEK/ERK signaling pathway and promotion of epithelial-mesenchymal transition (EMT) in cervical cancer cells. The interaction was identified by pulldown/MS and validated by bilateral co-immunoprecipitation.\",\n      \"method\": \"Pulldown assay, mass spectrometric peptide sequencing, bilateral co-immunoprecipitation, MEK/ERK phosphorylation assay, EMT marker analysis\",\n      \"journal\": \"Cancer letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — interaction identified by MS and confirmed by reciprocal Co-IP with functional downstream readout; single lab\",\n      \"pmids\": [\"25458953\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"HRAS is uniquely dependent on farnesyltransferase (FTase) for prenylation and membrane localization; NRAS and KRAS can use geranylgeranyl transferase as a bypass. Tipifarnib (FTase inhibitor) reduces HRAS processing and plasma membrane localization, decreasing GTP-bound HRAS and downstream effector pathway signaling (RAS effector pathways). This selectively inhibits growth of HRAS-mutant but not NRAS- or KRAS-mutant RMS xenografts in vivo.\",\n      \"method\": \"HRAS prenylation/processing assays, plasma membrane fractionation, GTP-bound HRAS (active Ras) pulldown, RAS effector pathway signaling (Western blot), 2D/3D cell growth assays, in vivo xenograft models with genomic RAS mutation stratification\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — direct biochemical demonstration of FTase-dependent HRAS processing and membrane localization with mutation-stratified in vivo validation; multiple orthogonal methods\",\n      \"pmids\": [\"35459782\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Oncogenic HRAS mutations (G12S, G12A) inhibit AMPK signaling, leading to impaired mitochondrial proteostasis and defective oxidative phosphorylation. This was demonstrated in CS mouse heart/skeletal muscle, patient fibroblasts, hiPSC-derived cardiomyocytes, and zebrafish models. Pharmacological activation of mitochondrial bioenergetics restored organelle function and reduced cardiac hypertrophy.\",\n      \"method\": \"CS mouse model (HrasG12S), patient skin fibroblasts, hiPSC-derived cardiomyocytes, HrasG12V zebrafish model, lentiviral HRAS mutant constructs, AMPK pathway analysis, mitochondrial function assays (respiration, proteostasis), pharmacological rescue\",\n      \"journal\": \"The Journal of clinical investigation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple independent model systems (mouse, human cells, fish, iPSC), convergent pathway (AMPK) identification, and pharmacological rescue; well-controlled study\",\n      \"pmids\": [\"35230976\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Inhibition of MEK/MAPK pathway signaling with a MEK inhibitor rescues HrasG12V-induced skeletal myopathy in a Costello syndrome mouse model. Activated HrasG12V causes skeletal myopathy via inhibition of embryonic myogenesis and myofiber formation, involving hyperactivation of Ras/MAPK and PI3K/AKT pathways and significant reduction in p38 signaling. MEK inhibition rescues the myopathy both in vitro and in vivo.\",\n      \"method\": \"HrasG12V knock-in mouse model, muscle histomorphometry, in vitro myogenesis assays, MEK inhibitor treatment in vitro and in vivo, phosphoprotein analysis (MAPK, AKT, p38), global transcriptomics\",\n      \"journal\": \"Disease models & mechanisms\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic mouse model with pharmacological rescue and multiple signaling readouts; single lab\",\n      \"pmids\": [\"34553752\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Mutant HRAS drives metastasis of head and neck cancer by suppressing the Hippo pathway and stabilizing YAP1, which transcriptionally activates AXL. Farnesyltransferase inhibitor tipifarnib activates the Hippo pathway, reduces nuclear YAP1 export, suppresses YAP1-mediated AXL expression, attenuates VEGFA/VEGFC expression, and reduces lymphovascular angiogenesis and metastasis. AXL depletion phenocopies tipifarnib in blocking invasion and metastasis.\",\n      \"method\": \"HRASmut human HNC cell lines, patient-derived xenografts, syngeneic mouse model, genetic (siRNA/shRNA) and pharmacological (tipifarnib) manipulation, targeted proteomics, Hippo pathway signaling analysis, in vivo metastasis assays\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo PDX and syngeneic models with genetic and pharmacological validation; single lab\",\n      \"pmids\": [\"36753744\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1987,\n      \"finding\": \"Oncogenic forms of H-ras (but not proto-oncogenic forms) completely suppress skeletal myoblast differentiation (fusion, nicotinic acetylcholine receptor and creatine kinase induction) at the level of muscle-specific mRNA accumulation, independent of effects on cell proliferation. This demonstrates H-ras mimics the differentiation-inhibitory effects of FGF and TGF-beta.\",\n      \"method\": \"DNA-mediated gene transfer into C2 myoblasts, myotube fusion assay, muscle-specific gene expression (Northern blot for AChR and creatine kinase mRNA), growth curve analysis\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — gain-of-function with oncogenic vs. proto-oncogenic forms, multiple differentiation readouts; foundational study, single lab\",\n      \"pmids\": [\"3600660\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"SPRED1 specifically perturbs K-ras (but not H-ras) membrane organization and ERK signaling at the plasma membrane by becoming enriched in acidic membrane domains. However, SPRED1 does block the positive effects of galectin-1 on H-ras nanoclustering. Legius syndrome-associated SPRED1 mutations show diminished binding to both galectin-1 and B-Raf.\",\n      \"method\": \"SPRED1 overexpression and membrane domain analysis, H-ras vs. K-ras ERK signaling assays, galectin-1 co-expression, Legius syndrome mutant analysis, plasma membrane fractionation\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct comparison of H-ras vs. K-ras with mechanistic membrane domain analysis and disease mutant validation; single lab\",\n      \"pmids\": [\"27503857\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"H-Ras and K-Ras exert distinct effects on apoptosis in endometrial cells: activated H-Ras rescues cells from apoptosis (dependent on PI3K/Akt), whereas activated K-Ras promotes apoptosis (dependent on Raf/MEK/MAPK). These isoform-specific differences depend on effector domain interactions, as demonstrated by K-Ras effector domain mutants (K12V35S preferentially activating MAPK/apoptosis; K12V40C preferentially activating Akt/survival).\",\n      \"method\": \"Stable expression of activated H-Ras and K-Ras, MEK inhibitor (U0126) and PI3K inhibitor (LY294002) epistasis, K-Ras effector domain mutants (35S and 40C), apoptosis assays, MAPK/Akt phosphorylation\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — effector domain mutant epistasis with isoform-specific inhibitor experiments; single lab\",\n      \"pmids\": [\"15087391\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"HRAS is a lipid-anchored small GTPase that cycles between GDP-bound (inactive) and GTP-bound (active) states, undergoes sequential post-translational modifications (farnesylation by FTase — uniquely obligate for HRAS unlike KRAS/NRAS — followed by proteolysis, methylation, and palmitoylation at Cys181/184), and signals from distinct plasma membrane nanoclusters, endosomes, Golgi, and ER compartments to activate downstream cascades including Raf/MEK/ERK and PI3K/Akt; its membrane localization, palmitoylation-cycle dynamics, and signaling output are regulated by redox modification of palmitoylation-site cysteines, beta-TrCP/Wnt-mediated ubiquitin-proteasomal degradation, QPCT-mediated stabilization, m6A epitranscriptomic control of translation, G-quadruplex/i-motif promoter structures (bound by MAZ, Sp1, and hnRNP A1), ERK-dependent promotion of Mst1/Mst2 heterodimer formation to suppress the Hippo pathway, Aurora A-scaffold enhancement of Raf-1 recruitment, and TRPML1-mediated cholesterol transport that maintains plasma membrane nanoclustering; oncogenic HRAS mutations additionally impair AMPK-mitochondrial homeostasis and drive metastasis via YAP1-AXL axis activation.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"HRAS is a lipid-anchored small GTPase that signals through the Raf/MEK/ERK and PI3K/Akt cascades to drive proliferation, transformation, and developmental signaling [#10, #31]. Its activity depends on a sequence of post-translational membrane-targeting events: FTase catalyzes farnesylation through a mechanism in which the catalytic zinc coordinates the CAAX cysteine as an ionized thiolate, and FTase substrate is bound synergistically with farnesyl pyrophosphate [#1]; HRAS is uniquely obligate for FTase, since unlike NRAS and KRAS it cannot bypass FTase via geranylgeranyltransferase, making HRAS-mutant tumors selectively sensitive to FTase inhibition [#25]. A dynamic palmitoylation cycle at Cys181/184 — installed by palmitoylation and reversed by a palmitoyl-protein thioesterase that recognizes only native, nucleotide-bound H-Ras [#0] — governs membrane distribution; oxidation of these cysteines under metabolic stress blocks palmitoylation, redistributes HRAS from plasma membrane to Golgi, and suppresses ERK signaling [#7]. HRAS signaling is spatially partitioned: it activates strongest from the plasma membrane, regulates the broadest transcriptional program from the ER, and engages TP53-dependent survival from the Golgi [#17], with plasma-membrane signaling requiring nanocluster organization maintained by TRPML1-dependent cholesterol transport [#15] and HRAS-specific endocytic recycling for Raf-1 activation [#4, #16]. On the membrane HRAS forms switch-II-mediated dimers abolished by the Y64A mutation [#8]. Downstream, HRAS-ERK signaling drives transformation through an Egr1→KLF5→cyclin D1 axis [#10], suppresses the Hippo pathway by promoting low-activity Mst1/Mst2 heterodimers [#11], and in mutant head-and-neck cancer stabilizes YAP1 to transcriptionally activate AXL and drive metastasis [#28]. HRAS protein abundance is set by beta-TrCP/Wnt-regulated ubiquitin-proteasomal degradation [#6], QPCT-mediated stabilization [#18], alternative-splicing suppression of p21 expression [#2], and m6A-dependent translational control of the HRAS 3' UTR via FTO and YTHDF1 [#19]. Germline activating HRAS mutations cause Costello syndrome: the p.Lys117Arg mutation accelerates nucleotide dissociation ~80-fold while retaining normal hydrolysis [#14], and oncogenic G12 mutations produce skeletal myopathy and cardiac hypertrophy through MEK/MAPK hyperactivation and AMPK-mitochondrial dysfunction [#26, #27].\",\n  \"teleology\": [\n    {\n      \"year\": 1987,\n      \"claim\": \"Established that oncogenic HRAS does more than drive proliferation — it actively blocks a differentiation program, defining HRAS as a determinant of cell fate.\",\n      \"evidence\": \"Gene transfer of oncogenic vs proto-oncogenic H-ras into C2 myoblasts with muscle-specific gene expression readouts\",\n      \"pmids\": [\"3600660\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular intermediates linking HRAS to suppression of muscle-specific transcription not defined\", \"Did not distinguish effector pathways responsible\"]\n    },\n    {\n      \"year\": 1989,\n      \"claim\": \"Showed that HRAS protein levels are constrained pre-translationally, identifying alternative splicing of an intronic exon as a negative control over p21 H-Ras expression and transforming potential.\",\n      \"evidence\": \"Gene reconstruction, S1 nuclease analysis, and intronic mutational scanning\",\n      \"pmids\": [\"2667764\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Trans-acting splicing factors not identified\", \"Physiological signals that modulate exon inclusion unknown\"]\n    },\n    {\n      \"year\": 1993,\n      \"claim\": \"Identified an enzyme that reverses HRAS palmitoylation, establishing that the palmitoylation state is dynamically regulated and conformation-dependent.\",\n      \"evidence\": \"Purification of a 37-kDa palmitoyl-protein thioesterase from bovine brain and in vitro assay on native, nucleotide-bound H-Ras\",\n      \"pmids\": [\"7901201\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo relevance to HRAS membrane cycling not tested\", \"Gene identity of the thioesterase not established here\"]\n    },\n    {\n      \"year\": 1995,\n      \"claim\": \"Placed RHAMM-dependent focal adhesion signaling downstream of HRAS, connecting HRAS transformation to adhesion turnover and metastasis.\",\n      \"evidence\": \"Dominant suppressor and antisense RHAMM in H-ras-transformed fibroblasts with tumorigenicity, metastasis, and FAK phosphorylation readouts\",\n      \"pmids\": [\"7541721\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct biochemical link between HRAS and RHAMM not shown\", \"Mechanism of FAK regulation downstream of RHAMM unresolved\"]\n    },\n    {\n      \"year\": 1998,\n      \"claim\": \"Defined the catalytic chemistry of HRAS prenylation, showing the CAAX cysteine binds FTase as a zinc-coordinated thiolate with synergistic FPP binding.\",\n      \"evidence\": \"Fluorescence binding, pH-dependent affinity, and Co2+-substituted FTase spectroscopy\",\n      \"pmids\": [\"9799520\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Does not address downstream proteolysis/methylation steps\", \"In-cell kinetics not measured\"]\n    },\n    {\n      \"year\": 2002,\n      \"claim\": \"Revealed isoform-specific spatial requirements, showing HRAS (unlike KRAS) requires endocytosis/recycling and PI3K for Raf-1 activation.\",\n      \"evidence\": \"Dominant-negative dynamin and Rab5 mutants with Raf-1 kinase and PC12 differentiation assays\",\n      \"pmids\": [\"12077341\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Identity of the signaling-competent endosomal compartment not fully resolved\", \"Mechanism linking recycling to Raf-1 activation unknown\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Connected HRAS-ERK output to a transcriptional transformation program and to isoform-specific apoptotic control via effector selection.\",\n      \"evidence\": \"siRNA epistasis at Egr1/KLF5 and effector-domain mutant analysis with inhibitor epistasis in two cell systems\",\n      \"pmids\": [\"15077182\", \"15087391\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Generality of Egr1→KLF5→cyclin D1 axis across tissues untested\", \"Structural basis of effector-domain preference not defined\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Mapped active HRAS through stages of clathrin-independent endocytosis and macropinocytosis, defining where compartmentalized ERK/Akt activation occurs.\",\n      \"evidence\": \"Live imaging with phosphoinositide reporters and Arf6/Rab5 mutants\",\n      \"pmids\": [\"18094044\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single-lab imaging study\", \"Functional consequences of each endosomal stage on transcription not assessed\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Showed a Costello syndrome mutation activates HRAS by a novel biochemical route — accelerated nucleotide dissociation — distinct from canonical hydrolysis-defective G12/G13 mutations.\",\n      \"evidence\": \"Nucleotide exchange/hydrolysis/GAP assays plus crystal structure of recombinant p.Lys117Arg\",\n      \"pmids\": [\"17979197\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Cellular signaling magnitude relative to G12 mutants not quantified here\", \"GEF dependence of the fast-cycling mutant unaddressed\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Defined ubiquitin-proteasomal degradation of HRAS by beta-TrCP and showed Wnt/Axin/APC signaling sets HRAS abundance, establishing Wnt-Ras crosstalk.\",\n      \"evidence\": \"Reciprocal Co-IP to the beta-TrCP WD40 domain, ubiquitylation assays, gain/loss-of-function, and in vivo Wnt3a injection\",\n      \"pmids\": [\"19240121\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Phosphodegron on HRAS not mapped\", \"Whether oncogenic mutants escape this degradation untested\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Identified endoglin and TGF-beta/ALK5 as transcriptional and signaling regulators of HRAS expression and transformation capacity.\",\n      \"evidence\": \"Promoter transactivation, Ras-GTP pulldown, MEK/ERK readouts, and NIH3T3 focus formation with ALK5 inhibitor epistasis\",\n      \"pmids\": [\"20884686\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct cis-elements mediating TGF-beta induction not mapped\", \"Single-lab study\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Connected metabolic redox state to HRAS localization by showing oxidation of palmitoylation-site cysteines blocks palmitoylation and reroutes HRAS off the plasma membrane.\",\n      \"evidence\": \"MALDI-TOF/TOF identification of oxidized Cys181/184, C181/184S rescue, fractionation, and ERK/apoptosis readouts\",\n      \"pmids\": [\"22085642\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Reversibility of oxidation in vivo not quantified\", \"Enzymes mediating re-reduction unidentified\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Defined a structural/transcriptional control layer at the HRAS promoter, with G-quadruplex repressors bound by MAZ/Sp1 and i-motifs unfolded by hnRNP A1 to activate transcription.\",\n      \"evidence\": \"ChIP, EMSA, promoter mutagenesis, FRET/CD, and RNAi in cells\",\n      \"pmids\": [\"21931711\", \"26674223\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"In vivo dynamics of quadruplex/i-motif folding not measured\", \"Quantitative contribution to endogenous HRAS expression incomplete\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Extended HRAS biology to intercellular and pathogen contexts, showing HRAS transfers between cells via tunneling nanotubes and is required for HCV receptor complex assembly.\",\n      \"evidence\": \"Optical-tweezer/FRAP imaging with lipidation mutants; proteomics, RNAi, and lateral diffusion plus HCV infection assays\",\n      \"pmids\": [\"23868059\", \"23498955\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Physiological frequency and significance of TNT transfer unknown\", \"How HRAS GTPase activity drives CD81 lateral diffusion unresolved\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Demonstrated HRAS dimerizes on membranes through a switch-II interface, defining a membrane-dependent oligomeric state relevant to effector activation.\",\n      \"evidence\": \"FCS, photon-counting histogram, single-molecule tracking, supported bilayer reconstitution, and Y64A mutagenesis\",\n      \"pmids\": [\"24516166\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional consequence of dimerization for Raf activation not directly tested\", \"Role of dimer interface in cells unconfirmed\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Identified CIP2A as a direct HRAS partner driving MEK/ERK activation and EMT, expanding the HRAS interactome in cancer.\",\n      \"evidence\": \"Pulldown/MS and bilateral Co-IP with EMT and signaling readouts\",\n      \"pmids\": [\"25458953\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Interaction interface not mapped\", \"Single cancer-cell context\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Resolved how HRAS signaling is terminated and sustained, showing endogenous HRAS is plasma-membrane/recycling-endosome localized and that EGFR endocytosis spatially separates EGFR-Grb2 from HRAS.\",\n      \"evidence\": \"CRISPR mVenus-HRas knock-in, live imaging, surface EGFR inactivation, and ERK/MEK time courses\",\n      \"pmids\": [\"26858456\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Generality across receptor systems untested\", \"Quantitative contribution of recycling-endosome pool to signaling unresolved\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Linked HRAS to Hippo suppression and defined membrane-organization regulators, showing ERK promotes inactive Mst1/Mst2 heterodimers and that TRPML1 cholesterol transport maintains HRAS nanoclusters.\",\n      \"evidence\": \"Co-IP and in vitro kinase comparison with Mst1 KO; MCOLN1 knockdown/inhibition with nanoclustering and ERK readouts\",\n      \"pmids\": [\"27238285\", \"30787043\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct biochemical step from ERK to heterodimer formation incompletely defined\", \"How cholesterol geometry enforces nanoclustering not structurally resolved\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Clarified isoform-specific membrane regulation by SPRED1/galectin-1, distinguishing HRAS from KRAS nanocluster control.\",\n      \"evidence\": \"SPRED1 membrane-domain analysis, galectin-1 co-expression, and Legius syndrome mutant binding\",\n      \"pmids\": [\"27503857\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism by which SPRED1 blocks galectin-1's positive effect on HRAS unresolved\", \"Single-lab study\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Identified Aurora A as a scaffolding partner enhancing HRAS-Raf-1 binding within a trimeric complex, adding a kinase-independent layer to HRAS-MAPK activation.\",\n      \"evidence\": \"Reciprocal Co-IP, domain-mapping pulldowns, and MAPK readouts with active/inactive HRAS\",\n      \"pmids\": [\"28177880\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Structural basis of the trimeric complex unknown\", \"In vivo relevance untested\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Established that HRAS signaling identity is location-encoded, with distinct kinase, interaction, and transcriptional outputs from plasma membrane, ER, and Golgi.\",\n      \"evidence\": \"Compartment-targeted HRAS constructs with integrated phospho/interaction/transcriptomics\",\n      \"pmids\": [\"30865897\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Endogenous compartmental flux not measured\", \"Targeted constructs may not recapitulate native localization stoichiometry\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Identified QPCT as a direct HRAS-stabilizing partner that attenuates ubiquitination and confers drug resistance, complementing the beta-TrCP degradation axis.\",\n      \"evidence\": \"Proteome microarray, Co-IP, colocalization, ubiquitination assay, and ERK readouts in renal cell carcinoma\",\n      \"pmids\": [\"31534544\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether QPCT competes with beta-TrCP directly unknown\", \"Interaction interface not mapped\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Defined the unique FTase-dependence of HRAS and validated FTase inhibition as a mutation-selective therapeutic strategy.\",\n      \"evidence\": \"Processing, membrane fractionation, active-HRAS pulldown, and mutation-stratified RMS xenografts with tipifarnib\",\n      \"pmids\": [\"35459782\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Resistance mechanisms to sustained FTase inhibition not addressed\", \"Effect on non-canonical HRAS compartments untested\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Linked oncogenic HRAS to metabolic/organismal pathology, showing G12 mutations impair AMPK signaling and mitochondrial proteostasis in Costello syndrome models.\",\n      \"evidence\": \"CS mouse, patient fibroblasts, hiPSC cardiomyocytes, and zebrafish with AMPK and mitochondrial assays and pharmacological rescue\",\n      \"pmids\": [\"35230976\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct biochemical link from HRAS to AMPK inhibition not defined\", \"Whether ERK pathway mediates the mitochondrial defect untested\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Established m6A epitranscriptomic control specific to the HRAS 3' UTR, where FTO and YTHDF1 regulate translational elongation and tumor phenotypes.\",\n      \"evidence\": \"m6A-seq, site-mutated luciferase reporters, FTO/YTHDF perturbations, and polysome profiling\",\n      \"pmids\": [\"36996116\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Upstream signals controlling HRAS 3' UTR methylation unknown\", \"Single-lab study\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Resolved the atomic structure of the HRAS promoter i-motif and defined a YAP1-AXL metastatic axis driven by mutant HRAS, deepening both transcriptional-control and oncogenic-output models.\",\n      \"evidence\": \"1.77 Å crystal structure of iHRAS; HRASmut HNC cell/PDX/syngeneic models with tipifarnib and AXL depletion\",\n      \"pmids\": [\"36995904\", \"36753744\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"In-cell relevance of the crystallographic i-motif fold not confirmed\", \"Mechanism of YAP1 stabilization by mutant HRAS incompletely defined\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How the multiple layers of HRAS abundance control (splicing, beta-TrCP degradation, QPCT stabilization, m6A translation, promoter quadruplex/i-motif structures) are integrated and coordinated with compartment-specific signaling in normal vs. oncogenic cells remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No unified model linking expression-level control to spatial signaling output\", \"Crosstalk among the regulatory layers untested in a single system\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0003924\", \"supporting_discovery_ids\": [14, 25, 9]},\n      {\"term_id\": \"GO:0060089\", \"supporting_discovery_ids\": [10, 17, 31]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [7, 16, 15, 8]},\n      {\"term_id\": \"GO:0005768\", \"supporting_discovery_ids\": [4, 5, 16]},\n      {\"term_id\": \"GO:0005794\", \"supporting_discovery_ids\": [7, 17]},\n      {\"term_id\": \"GO:0005783\", \"supporting_discovery_ids\": [17]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [4, 10, 11, 17]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [14, 26, 27, 28]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [1, 6, 18, 25]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"RAF1\", \"AURKA\", \"CIP2A\", \"QPCT\", \"BTRC\", \"CD81\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":8,"faith_total":8,"faith_pct":100.0}}