| 1996 |
RIT1 (Rit) and Rin define a novel subfamily of Ras-related proteins. Rit binds GTP in vitro, is ubiquitously expressed, and localizes to the plasma membrane despite lacking classical C-terminal lipidation signals; a C-terminal cluster of basic amino acids is important for Rit membrane binding (deletion analysis). |
Degenerate PCR cloning, GTP-binding assay in vitro, deletion analysis, subcellular localization by expression in cells |
The Journal of Neuroscience |
Medium |
8824319
|
| 1999 |
RIT1 (Rit) and Rin exhibit intrinsic GTPase activity; conversion of Gln79 to Leu (equivalent to Ras position 61) abolishes GTPase activity. Both proteins display higher GTP dissociation rates (5–10× faster) than most Ras-like GTPases. Yeast two-hybrid analysis showed Rit and Rin interact with RalGDS, Rlf, and AF-6/Canoe in a GTP- and effector domain-dependent manner, but not with Raf kinases, RIN1, or the p110 subunit of PI3K. |
Recombinant protein biochemical characterization (GTPase assay, nucleotide dissociation kinetics), active-site mutagenesis (Q79L), yeast two-hybrid |
Archives of Biochemistry and Biophysics |
High |
10545207
|
| 2000 |
Constitutively active Rit (Rit79L) transforms NIH3T3 cells (soft agar, tumor formation in nude mice) without activating ERK, JNK, p38 MAPK, or PI3K/Akt in immune complex kinase assays. Rit cooperates with Raf and RhoA to form foci. Rit-transformed cell survival depends on a farnesylated protein (farnesyltransferase inhibitors caused apoptosis), despite Rit itself lacking lipidation signals. |
Stable NIH3T3 expression of constitutively active Rit, soft agar assay, nude mouse tumorigenesis, immune complex kinase assays, focus assays, farnesyltransferase inhibitor treatment |
Oncogene |
High |
11032018
|
| 2000 |
RGL3, a novel RalGEF-like protein, interacts with RIT1 (Rit) in a GTP- and effector loop-dependent manner via its C-terminal 99-amino acid domain. RGL3 exhibits guanine nucleotide exchange activity toward the small GTPase Ral, and this activity is stimulated in vivo by activated Rit or Ras, identifying RGL3 as a Rit downstream effector. |
Yeast two-hybrid screen, in vitro GEF assay for Ral, in vivo co-expression of activated Rit/Ras with RGL3 |
The Journal of Biological Chemistry |
High |
10869344
|
| 2002 |
Constitutively active Rit (RitL79) in PC6 pheochromocytoma cells induces neuronal differentiation (neurite outgrowth) and promotes cell survival against growth factor withdrawal. Rit-mediated effects on neurite outgrowth and survival were blocked by dominant-negative C-Raf1 or MEK1 co-expression. Active Rit stimulated ERK1/2 phosphorylation in PC6 cells but not COS cells, indicating cell-type specificity. Survival did not involve PI3K/Akt. |
Constitutively active mutant expression in PC6 cells, dominant-negative co-expression, Western blot for ERK phosphorylation, pharmacological MEK inhibition, apoptosis assay |
The Journal of Biological Chemistry |
High |
11914372
|
| 2002 |
Constitutively active Rit stimulates p38γ but not p38α, p38δ, ERK5, or classical MAPK in NIH3T3 cells, and p38γ activation is required for Rit-stimulated gene expression and cellular transformation. |
Overexpression of activated Rit mutants, kinase activity assays for individual p38 isoforms and ERK5, luciferase reporter assays |
FEBS Letters |
Medium |
11821041
|
| 2003 |
In human neuroblastoma SH-SY5Y cells, constitutively active Rit increases neurite initiation, elongation, and branching. MEK inhibition blocked Rit-induced neurite initiation but not elongation or branching, demonstrating both MEK-dependent and MEK-independent signaling mechanisms downstream of Rit. |
Adenoviral expression of Rit mutants, image analysis of neurite morphology, MEK inhibitor (PD098059), Western blot for ERK1/2 and Akt |
Journal of Cell Science |
Medium |
12668729
|
| 2005 |
NGF stimulation rapidly and sustainably activates Rit in PC6 cells. Active Rit promotes neuronal differentiation via B-Raf (not C-Raf) activation leading to sustained ERK and p38 MAPK signaling. Rit-induced differentiation is dependent on both MEK/ERK and p38 cascades. Rit knockdown by siRNA significantly attenuated NGF-dependent p38 and ERK activation and neuronal differentiation. |
Rit activation assay (GTP-loading), B-Raf/C-Raf binding assay, MEK and p38 pharmacological inhibitors, Rit siRNA knockdown in PC6 cells, neuronal differentiation assay |
Molecular and Cellular Biology |
High |
15632082
|
| 2005 |
Rin and Rit directly bind to the PDZ domain of PAR6 (cell polarity protein) in a GTP-dependent manner both in vivo and in vitro. Rin/Rit can form a ternary complex with PAR6 and Rac/Cdc42 that synergistically potentiates cell transformation in NIH3T3 cells; the Rin/Rit–PDZ interaction is required for this effect. |
Co-immunoprecipitation (in vivo), in vitro binding assay with purified proteins, NIH3T3 transformation/focus assay |
The Journal of Biological Chemistry |
Medium |
15831491
|
| 2006 |
PACAP38 activates Rit in a cAMP-dependent but PKA-independent manner downstream of the Epac guanine nucleotide exchange factor. Rit knockdown (RNAi) selectively suppressed PACAP38-elicited p38 MAPK activation (but not ERK) and inhibited PACAP38-mediated CREB-dependent transcription and neurite outgrowth. Epac is required for PACAP38-mediated Rit activation, but neither Epac1 nor Epac2 activates Rit directly, indicating a novel indirect mechanism not involving Rap GTPases. |
Rit activation assay, RNAi knockdown, pharmacological inhibition (PKA inhibitors), p38/ERK Western blot, CREB luciferase reporter, neurite outgrowth assay |
Molecular and Cellular Biology |
High |
17000774
|
| 2007 |
Dominant-negative Rit in hippocampal neurons inhibits axonal growth but potentiates dendritic growth; constitutively active Rit promotes axonal growth but inhibits dendritic growth. BMP7 treatment that promotes dendritic growth decreases Rit-GTP loading. Pharmacological MEK1 inhibition blocks the axon-promoting and dendrite-inhibiting effects of active Rit, placing Rit upstream of ERK1/2 in differential axonal versus dendritic regulation. |
Dominant-negative and constitutively active Rit expression in primary hippocampal and sympathetic neurons, GTP-loading assay, MEK inhibitor, morphometric analysis |
The Journal of Neuroscience |
High |
17460085
|
| 2007 |
Rit effector loop mutant analysis demonstrated that Rit-mediated neuritogenesis requires MEK/ERK signaling but is independent of RalGEF activation. A novel mechanism of Par6 interaction was identified in vivo binding studies, suggesting Par6 may spatially restrict Rit signaling. |
Effector loop mutagenesis of Rit, in vivo binding assays (Co-IP), MEK inhibitor-based dissection of neuritogenesis |
Biochimica et Biophysica Acta |
Medium |
17976838
|
| 2008 |
IFNγ activates Rit (increased GTP loading) in pheochromocytoma cells and hippocampal neurons. Dominant-negative Rit or Rit RNAi suppressed IFNγ-induced p38 MAPK activation and dendritic retraction in sympathetic and hippocampal neurons. Pharmacological p38 inhibition blocked the dendrite-retracting effect without affecting STAT1 activation, identifying a novel IFNγ–Rit–p38 pathway controlling dendritic retraction. |
GTP-loading assay, dominant-negative Rit expression, RNAi knockdown, p38 inhibitor, dendritic morphometry, STAT1 Western blot |
Journal of Neurochemistry |
High |
18957053
|
| 2008 |
Rit (Goalpha-interacting protein identified by co-IP): the α-subunit of Go (Goalpha) interacts with Rit. Dominant-negative Rit inhibited Goalpha-induced neurite outgrowth and ERK phosphorylation in Neuro2a cells, establishing Rit as a downstream effector of Goalpha-mediated neuronal differentiation. |
Co-immunoprecipitation, dominant-negative Rit expression, ERK phosphorylation Western blot, neurite outgrowth assay |
Neuroreport |
Medium |
18388731
|
| 2010 |
PACAP-mediated Rit activation involves Src family kinase-dependent TrkA receptor transactivation. PACR1 stimulation triggers Gi-alpha and Gs-alpha/cAMP/Epac cascades leading to Src kinase activity and TrkA tyrosine phosphorylation. Src inhibition or lack of functional Trk receptors inhibits PACAP-mediated Rit activation. TrkA Y499 phosphorylation is critical for both PACAP-mediated transactivation and Rit activation. RNAi silencing of SOS1/2 inhibited Rit activation, implicating a TrkA/Shc/SOS complex in Rit regulation. |
Rit activation assay, Src inhibitor, TrkA mutants, phospho-Western blot, RNAi knockdown of SOS1/2 |
Molecular Biology of the Cell |
High |
20219970
|
| 2011 |
Rit promotes cell survival by directing a p38 MAPK-dependent Akt activation pathway. Rit shRNAi-treated cells show increased apoptosis and disrupted p38 signaling after stress. Constitutively active Rit promotes p38-Akt-dependent survival. Rit (but not Ras or Rap) associates with and is required for stress-mediated activation of the scaffolded p38-MK2-HSP27-Akt pro-survival complex. |
shRNAi knockdown, constitutively active Rit expression, Co-IP (Rit with p38/MK2/HSP27/Akt), apoptosis assay, Western blot for signaling components |
Molecular and Cellular Biology |
High |
21444726
|
| 2011 |
Rit GTPase is required for an evolutionarily conserved p38 MAPK-dependent oxidative stress survival pathway. Rit knockout mouse embryonic fibroblasts display increased apoptosis and selective disruption of p38-MK2-HSP27 signaling and downstream Akt activation (directing Bad phosphorylation) upon ROS exposure, but not after ER stress or DNA damage. Drosophila D-RIC null flies show increased susceptibility to environmental stresses and reduced p38 signaling, extending the pathway to invertebrates. |
Rit knockout mice (MEFs), Drosophila D-RIC null mutants, apoptosis assays, Western blot (p38/MK2/HSP27/Akt/Bad phosphorylation) |
Molecular Biology of the Cell |
High |
21737674
|
| 2012 |
Rit-mediated stress resistance requires a p38-MSK1/2-CREB activation cascade. Rit shRNAi silencing, p38 inhibition, or MSK1/2 inhibition disrupts stress-mediated CREB-dependent transcription and increases cell death. Active Rit stimulates CREB-Ser133 phosphorylation and induces Bcl-2 and Bcl-XL expression to promote survival. |
RNAi knockdown of Rit, pharmacological inhibition of p38 and MSK1/2, constitutively active Rit expression, CREB-Ser133 phospho-Western, luciferase reporter for CREB-dependent transcription, Bcl-2/Bcl-XL Western blot |
The Journal of Biological Chemistry |
Medium |
23038261
|
| 2012 |
Rit GTPase promotes survival of immature (Dcx+) hippocampal neurons but not neural progenitors (Nestin+) following oxidative stress. Rit-/- mice exhibit greater loss of adult-born immature neurons and blunted remodeling after traumatic brain injury. Rit-/- hippocampal cultures show blunted MAPK cascade activation after oxidative stress without affecting BDNF-dependent signaling. |
Rit knockout mice, hippocampal cultures, TBI model, immunofluorescence (Dcx/Nestin markers), Western blot for MAPK activation, apoptosis assay |
The Journal of Neuroscience |
High |
22815504
|
| 2012 |
Transgenic expression of constitutively active Rit (Q79L) selectively in neurons (Synapsin I promoter) confers dramatic oxidative stress resistance in hippocampal neurons. Pharmacological studies demonstrated p38 MAPK (not MEK/ERK) is required for Rit-mediated neuronal protection. |
Transgenic mouse overexpressing active Rit in neurons, pharmacological p38 and MEK inhibitors, oxidative stress challenge of hippocampal neurons |
Neuroscience Letters |
Medium |
23123784
|
| 2013 |
Gain-of-function missense mutations in RIT1 cause Noonan syndrome. Five RIT1 mutations identified in patients enhanced ELK1 transactivation in luciferase assays in NIH 3T3 cells. Introduction of mutant RIT1 mRNAs into zebrafish embryos caused craniofacial abnormalities, cardiac looping defects, hypoplastic heart chambers, and elongated yolk sac—phenotypes consistent with other RASopathy mutations. |
Exome sequencing (discovery), Sanger sequencing (validation), ELK1 luciferase reporter assay in NIH 3T3 cells, zebrafish mRNA injection model |
American Journal of Human Genetics |
High |
23791108
|
| 2014 |
Somatic RIT1 mutations clustering near the switch II domain occur in ~2% of lung adenocarcinomas and are mutually exclusive with all other known driver mutations. Ectopic expression of mutant RIT1 induces cellular transformation in vitro and tumor formation in vivo (xenografts), reversible by combined PI3K and MEK inhibition. |
Cancer genome sequencing (discovery), ectopic expression in cell lines (transformation assay), xenograft in vivo models, combined PI3K+MEK inhibitor treatment |
Oncogene |
High |
24469055
|
| 2014 |
mTORC2 is a critical downstream mediator of Rit-dependent survival signaling in response to ROS. Rit physically interacts with Sin1 (MAPKAP1, an mTORC2 component). Rit loss compromises ROS-dependent mTORC2 complex activation and blunts mTORC2-mediated phosphorylation of Akt. |
Co-immunoprecipitation (Rit–Sin1 interaction), ROS treatment, mTORC2 activity assay, Akt phosphorylation Western blot in Rit-/- cells |
PloS One |
Medium |
25531880
|
| 2016 |
RIT1 disease-associated mutations biochemically fall into two classes: mutations S35T, A57G, and Y89H exhibit accelerated nucleotide exchange (increased GTP loading), while F82V and T83P impair GTP hydrolysis. RIT1 intrinsic nucleotide exchange rate is ~4-fold faster than H-RAS. All disease-associated mutations increase the GTP-loaded activated state of RIT1 in vitro and in cells. |
Real-time NMR-based GTPase assay for RIT1 and disease-associated mutants, Ras-binding domain pulldown assay in HEK293T cells |
The Journal of Biological Chemistry |
High |
27226556
|
| 2016 |
RIT1 controls Sox2 transcriptional activity in hippocampal neuronal precursor cells via an Akt-dependent signaling cascade, leading to Sox2 stabilization and transcriptional activation. RIT1 loss blunts Akt signaling and neurogenesis. In vivo, Sox2-dependent hippocampal neurogenesis is significantly impaired after IGF-1 infusion in RIT1-/- mice. |
Hippocampal neuronal precursor cell culture, RIT1-/- knockout mice, Sox2 phosphorylation Western blot, Akt inhibition, gene expression profiling, in vivo IGF-1 infusion |
The Journal of Biological Chemistry |
Medium |
28007959
|
| 2017 |
IGF-1 stimulates a RIT1-dependent increase in Sox2 levels in hippocampal neuronal precursor cells, resulting in pro-neural gene expression and cellular proliferation. RIT1 stimulates Akt-dependent phosphorylation of Sox2 at T118, leading to its stabilization and transcriptional activation. RIT1-/- HNPCs show deficient IGF-1-dependent Akt signaling and neuronal differentiation. Exercise-mediated potentiation of hippocampal neurogenesis is diminished in RIT1-/- mice. |
Hippocampal neuronal precursor cell culture, RIT1-/- knockout mice, Sox2-T118 phospho-specific Western blot, Akt inhibition, in vivo IGF-1 infusion, running exercise model |
Scientific Reports |
High |
28607354
|
| 2018 |
RIT1 directly interacts with PAK1 (identified as a novel direct effector), and also directly interacts with the Rho GTPases CDC42 and RAC1; these interactions are independent of the guanine nucleotide bound to RIT1. Disease-causing RIT1 mutations enhance protein-protein interactions with PAK1, CDC42, and RAC1 and uncouple complex formation from serum/growth factor dependence. The RIT1-PAK1 complex regulates cytoskeletal rearrangements (dissolution of stress fibers, reduction of focal adhesions) via CDC42/RAC1 and kinase-active PAK1. RIT1 wildtype and disease variants enhance cell motility. |
Heterologous expression, pulldown with purified recombinant proteins, Co-IP, stress fiber/focal adhesion imaging in COS7 cells, transwell migration assay, dominant-negative CDC42/RAC1 and kinase-dead PAK1 rescue experiments |
PLoS Genetics |
High |
29734338
|
| 2019 |
LZTR1 acts as a substrate adaptor for a Cullin3-RING E3 ubiquitin ligase complex that binds, ubiquitinates, and promotes proteasomal degradation of RIT1. Pathogenic mutations in either RIT1 (near the switch II domain) or LZTR1 result in incomplete degradation of RIT1, leading to RIT1 protein accumulation and dysregulated growth factor signaling. A knock-in mouse model with RIT1 mutation develops a Noonan syndrome-like phenotype. |
Mass spectrometry (identified LZTR1 as RIT1 interactor), Co-IP, proteasome inhibitor experiments, isogenic germline knock-in mouse model, Western blot for RIT1 protein levels |
Science |
High |
30872527
|
| 2019 |
Rit1 A57G knock-in mice (Noonan syndrome model) exhibit cardiac hypertrophy, cardiac fibrosis (with increased S100A4, vimentin, periostin), and other NS-associated features. Biochemical analysis shows elevated phospho-Akt (Thr308) in embryos and in isoproterenol-treated Rit1A57G/+ hearts, implicating the AKT signaling pathway in RIT1 A57G mutant downstream signaling. |
Knock-in mouse generation (Rit1-A57G), echocardiography, histopathology, isoproterenol stimulation, Western blot for phospho-Akt |
EBioMedicine |
Medium |
30898653
|
| 2021 |
RIT1 is essential for timely mitotic progression and proper chromosome segregation. During mitosis, RIT1 dissociates from the plasma membrane and interacts directly with spindle assembly checkpoint (SAC) proteins MAD2 and p31comet in a CDK1 activity-dependent manner. Pathogenic RIT1 silences the SAC by sequestering MAD2 from the mitotic checkpoint complex (MCC), accelerating mitotic transit and promoting chromosome segregation errors and aneuploidy. |
Live-cell imaging, immunofluorescence, Co-IP of RIT1 with MAD2/p31comet, CDK1 inhibitor experiments, chromosome segregation and aneuploidy assays, SAC functional assays |
Current Biology |
High |
34237269
|
| 2021 |
Multiomic profiling showed that mutant RIT1 and mutant KRAS both promote canonical RAS signaling (RAF/MEK/ERK) and that overexpression of wild-type RIT1 partially phenocopies oncogenic RIT1 and KRAS, including induction of epithelial-to-mesenchymal transition, indicating RIT1 protein abundance is a factor in its pathogenic function. |
Quantitative proteomics, phosphoproteomics, and transcriptomics in isogenic lung epithelial cells expressing WT and mutant RIT1 or KRAS |
Science Signaling |
Medium |
34846918
|
| 2022 |
Cross-species analysis in Drosophila and mice demonstrated that LZTR1 preferentially ubiquitinates and promotes degradation of RIT1 orthologs over classical RAS GTPases. Embryonic lethality of homozygous Lztr1 null mice is rescued by co-deletion of Rit1, genetically establishing RIT1 as the primary in vivo substrate of LZTR1 in mice. |
Drosophila Lztr1 loss-of-function, mouse Lztr1 knockout and Lztr1/Rit1 double knockout, ubiquitination assays, Western blot for RIT1 orthologue protein levels |
eLife |
High |
35467524
|
| 2023 |
RAF kinases are direct effectors of membrane-bound mutant RIT1 required for MAPK activation. RIT1 contains critical residues that facilitate interaction with membrane lipids, and these are necessary for RIT1 association with RAF kinases and MAPK activation. Although mutant RIT1 binds RAF kinases directly, it fails to activate MAPK signaling in the absence of classical RAS proteins. Pathway inhibition (targeting RAS-RAF-MAPK) alleviates cardiac hypertrophy in a RIT1 mutant Noonan syndrome mouse model. |
RAF kinase co-immunoprecipitation, lipid-binding mutagenesis, MAPK signaling assays in cells with/without RAS depletion, Noonan syndrome knock-in mouse cardiac phenotype rescue with pathway inhibitor |
Science Advances |
High |
37450595
|
| 2023 |
RIT1 interacts with SMC3 and PDS5 during mitosis (Co-IP + mass spectrometry). RIT1 protects and maintains SMC3 acetylation by binding to SMC3 and PDS5, thereby promoting cell division and proliferation in hepatocellular carcinoma. RIT1 knockdown disrupts mitosis, causes G2/M phase arrest, mitotic catastrophe, and apoptosis. |
Co-immunoprecipitation, mass spectrometry, RNA-seq, Western blot for SMC3 acetylation, immunofluorescence, flow cytometry (cell cycle), live cell imaging, in vivo xenograft |
Journal of Experimental & Clinical Cancer Research |
Medium |
38017479
|
| 2024 |
USP9X deubiquitinase stabilizes both wild-type and mutant RIT1 protein. Depletion of USP9X decreases RIT1 protein stability and abundance, and resensitizes RIT1-mutant cells to EGFR tyrosine kinase inhibitors in vitro and in vivo. |
USP9X knockdown/depletion, RIT1 protein stability assays, ubiquitination assay (USP9X as deubiquitinase of RIT1), EGFR inhibitor sensitivity assays in vitro and in vivo |
iScience |
Medium |
39161959
|
| 2025 |
Physiologic expression of RIT1 M90I is sufficient to drive autochthonous lung tumor development in vivo in mouse models. RIT1 M90I tumors are sensitive to SHP2 inhibitors and RAS nucleotide exchange inhibition. RAS tri-complex inhibitors bind directly to GTP-bound RIT1 and cause tumor shrinkage. |
Knock-in/transgenic mouse autochthonous lung tumor models, SHP2 inhibitor treatment, RAS nucleotide exchange inhibitor, chemical biology/RAS tri-complex inhibitor direct binding to GTP-bound RIT1, tumor imaging |
Cancer Research |
High |
40644578
|
| 2021 |
The C-terminal peptide (CTP) of RIT1 associates with anionic lipid bilayers containing phosphatidylserine via electrostatic (charge complementarity) interactions. Molecular dynamics simulations showed the CTP is unstructured and that a 12-residue region binds strongly to anionic bilayers, providing the membrane-anchoring mechanism in the absence of prenylation. |
Molecular dynamics simulations of RIT1 CTP with lipid bilayers of varying composition |
Computational Biology and Chemistry |
Low |
33517146
|