{"gene":"RASSF2","run_date":"2026-06-10T06:43:36","timeline":{"discoveries":[{"year":2003,"finding":"RASSF2 binds directly to K-Ras in a GTP-dependent manner via the Ras effector domain, but only weakly interacts with H-Ras, establishing it as a K-Ras-specific effector. RASSF2 promotes apoptosis and cell cycle arrest.","method":"Direct binding assay (pulldown), GTP-dependence assay, cell-based apoptosis and cell cycle assays","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal binding assay with GTP-dependence, replicated in subsequent studies across multiple labs","pmids":["12732644"],"is_preprint":false},{"year":2005,"finding":"RASSF2 induces morphological changes and apoptosis when ectopically expressed in colorectal cancer cells, and inactivation of RASSF2 enhances K-Ras-induced oncogenic transformation, placing RASSF2 downstream of K-Ras as a negative regulator of transformation.","method":"Colony formation assay, flow cytometry, immunofluorescence microscopy, functional complementation","journal":"Gastroenterology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal cell-based assays in a single lab","pmids":["16012945"],"is_preprint":false},{"year":2007,"finding":"RASSF2 contains a functional bipartite nuclear localization signal (NLS); mutation of the NLS abolishes nuclear localization and significantly diminishes its tumor suppressor (growth suppression) activity, demonstrating that nuclear localization is required for full RASSF2 function.","method":"Site-directed mutagenesis of NLS, immunofluorescence localization, in vitro and in vivo growth suppression assays","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 2 / Strong — mutagenesis combined with localization and functional growth suppression assays in vitro and in vivo","pmids":["17891178"],"is_preprint":false},{"year":2009,"finding":"RASSF2 associates with the proapoptotic kinases MST1 and MST2 via its SARAH domain, and this interaction is confirmed at endogenous levels. RASSF2 co-immunoprecipitates active MST1/2, is phosphorylated by a co-immunoprecipitating kinase (likely MST1/2), and stabilizes MST2 by protecting it from proteolytic degradation. RASSF2 alone localizes to the nucleus, but co-expression with MST1 or MST2 results in cytoplasmic co-localization.","method":"Co-immunoprecipitation (endogenous), immunofluorescence co-localization, stable/transient expression, RASSF2 knockdown, kinase activity assay","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 2 / Strong — endogenous Co-IP, multiple orthogonal methods, knockdown and overexpression, replicated in subsequent studies","pmids":["19525978"],"is_preprint":false},{"year":2009,"finding":"MST1 regulates RASSF2 protein stability: knockdown of MST1 markedly destabilizes RASSF2, and Mst1-deficient mice show reduced Rassf2 protein levels. Conversely, RASSF2 activates MST1 kinase activity through formation of a RASSF2-MST1 complex. RASSF2 also activates JNK signaling and induces apoptosis in an MST1-independent manner.","method":"siRNA knockdown, Mst1 knockout mice, kinase activity assay, complex formation (Co-IP), apoptosis assay, JNK pathway assay","journal":"Biochemical and biophysical research communications","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic (knockout mice) plus biochemical (kinase assay, Co-IP) evidence, multiple orthogonal methods","pmids":["19962960"],"is_preprint":false},{"year":2009,"finding":"RASSF2 shuttles between nucleus and cytoplasm via a CRM-1-dependent nuclear export mechanism. A functional nuclear export signal (NES) resides in amino acids 240–260 (C-terminus); mutation of conserved residues Ile254, Val257, and Leu259 impairs nuclear export and interaction with CRM-1. ERK2 (MAPK) phosphorylates RASSF2 and this phosphorylation is required for nuclear export; inhibition of MAPK pathway blocks RASSF2 export. Nuclear import-defective RASSF2 fails to induce G1/S cell cycle arrest or apoptosis.","method":"NES mutagenesis, leptomycin B treatment, CRM-1 interaction assay, MAPK inhibitor treatment, in vitro phosphorylation assay, cell cycle and apoptosis assays","journal":"Experimental cell research","confidence":"High","confidence_rationale":"Tier 1-2 / Moderate — mutagenesis, pharmacological inhibition, in vitro phosphorylation, and functional phenotype in one study","pmids":["19555684"],"is_preprint":false},{"year":2010,"finding":"RASSF2 forms a direct and endogenous complex with PAR-4 (prostate apoptosis response protein 4). This interaction is regulated by K-Ras and is essential for the full apoptotic effects of PAR-4. RASSF2 (a primarily nuclear protein) modulates nuclear translocation of PAR-4 from cytoplasm to nucleus in prostate tumor cells, providing a mechanism for PAR-4's biological apoptotic effects.","method":"Co-immunoprecipitation (endogenous), subcellular fractionation/localization, functional apoptosis assay","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 2 / Moderate — endogenous Co-IP, localization experiments, and functional apoptosis assay in one rigorous study","pmids":["20368356"],"is_preprint":false},{"year":2010,"finding":"Deletion of the MST interaction domain of RASSF2 significantly reduces RASSF2-induced apoptosis in thyroid cancer cells, establishing that the MST interaction domain is functionally required for RASSF2-mediated apoptosis.","method":"Domain deletion mutagenesis, apoptosis assay in thyroid cancer cells","journal":"Molecular cancer","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — deletion mutagenesis with quantitative apoptosis readout in a single lab","pmids":["20920251"],"is_preprint":false},{"year":2008,"finding":"A RASSF2 deletion mutant lacking the Ras-association domain, unable to interact with Ras, exhibits less pro-apoptotic activity than the full-length protein, demonstrating that the pro-apoptotic activity of RASSF2 is related to its association with Ras.","method":"Deletion mutagenesis, apoptosis assay in OSCC cells","journal":"Cancer science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — domain deletion mutagenesis combined with functional apoptosis assay, single lab","pmids":["18294275"],"is_preprint":false},{"year":2012,"finding":"Loss of RASSF2 expression in lung cancer cells correlates with increased levels of activated AKT, indicating RASSF2 modulates Ras/AKT signaling. RASSF2 and K-Ras form an endogenous complex (confirmed by Co-IP), validating RASSF2 as a bona fide K-Ras effector in lung cancer cells.","method":"RNAi knockdown, Co-immunoprecipitation (endogenous), AKT phosphorylation assay, cell proliferation/invasion/anchorage-independent growth assays","journal":"Molecular biology international","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — endogenous Co-IP and signaling assay, single lab with multiple functional readouts","pmids":["22693671"],"is_preprint":false},{"year":2012,"finding":"Rassf2 knockout mice develop bone remodeling defects and haematopoietic anomalies. RASSF2 ablation suppresses osteoblastogenesis and promotes osteoclastogenesis in vitro. RASSF2 associates with IKKα and IKKβ and suppresses IKK kinase activity, thereby restraining NF-κB signaling during osteoclast and osteoblast differentiation. Reintroduction of RASSF2 or dominant-negative IKK normalizes differentiation in Rassf2-/- precursors.","method":"Knockout mouse model, bone marrow transplantation, in vitro differentiation assay, Co-immunoprecipitation (RASSF2-IKKα/β), IKK kinase activity assay, dominant-negative rescue","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — genetic knockout model, Co-IP, kinase assay, rescue experiment, multiple orthogonal methods in one rigorous study","pmids":["22227519"],"is_preprint":false},{"year":2016,"finding":"Proteomics-based screening identified novel RASSF2 interaction partners including C1QBP and Vimentin. The RASSF2-C1QBP interaction is enhanced by K-Ras, while the RASSF2-Vimentin interaction is reduced by K-Ras. RASSF2/K-Ras regulates the acetylation of Vimentin.","method":"Proteomics/MS pulldown, Co-immunoprecipitation validation, acetylation assay","journal":"Cancers","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — proteomics screen validated by Co-IP for two interactions and acetylation assay, single lab","pmids":["26999212"],"is_preprint":false},{"year":2017,"finding":"RASSF2 is a direct target of miR-7 in cancer-associated fibroblasts. Overexpression of miR-7 down-regulates RASSF2, which reduces PAR-4 secretion from fibroblasts and enhances proliferation and migration of co-cultured cancer cells, placing RASSF2 in a miR-7-RASSF2-PAR-4 signaling axis.","method":"miRNA overexpression/inhibition, co-culture assay, bioinformatic target validation, functional proliferation/migration assay","journal":"Oncotarget","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — functional miRNA/target assays with co-culture readout, single lab","pmids":["27901488"],"is_preprint":false},{"year":2020,"finding":"RASSF2 suppresses t(8;21) AML development, and its function depends on interaction with Hippo kinases MST1 and MST2, but is independent of canonical Hippo pathway signaling. Proximity-based biotin labeling revealed RASSF2 associates with Rac GTPase-related proteins including the guanine nucleotide exchange factor DOCK2. RASSF2 knockdown impairs Rac GTPase activation.","method":"Re-expression in AML models (in vitro and in vivo), proximity-based biotin labeling (BioID), Co-immunoprecipitation, Rac GTPase activation assay, MST1/2 interaction studies","journal":"Blood cancer journal","confidence":"High","confidence_rationale":"Tier 2 / Strong — BioID proteomics, Co-IP, functional rescue in multiple AML models, Rac activation assay, multiple orthogonal methods","pmids":["32029705"],"is_preprint":false},{"year":2019,"finding":"The miR-200 family directly targets RASSF2 mRNA; knockdown of the miR-200 family in HT-29 colon cancer cells increases RASSF2 expression and decreases ERK phosphorylation (MAPK/ERK signaling), while increased miR-200 expression decreases RASSF2 and increases ERK phosphorylation.","method":"siRNA knockdown, miRNA gain/loss of function, western blot for ERK phosphorylation, cell proliferation assay","journal":"Oncology letters","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — direct miRNA-target functional experiments with signaling readout, single lab","pmids":["31565080"],"is_preprint":false},{"year":2023,"finding":"RASSF2 mediates oxidative stress-induced apoptosis in lens epithelial cells by regulating AKT (Ser473) phosphorylation. Overexpression of RASSF2 promotes apoptosis under oxidative conditions, while knockdown reduces it. EGCG inhibits this pathway by reducing RASSF2 levels, thereby suppressing AKT phosphorylation and apoptosis.","method":"Overexpression and siRNA knockdown in lens epithelial cells, western blot for AKT Ser473 phosphorylation, mouse model","journal":"European journal of pharmacology","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — overexpression/knockdown with phosphorylation readout, in vitro and in vivo, single lab","pmids":["37979829"],"is_preprint":false},{"year":2026,"finding":"Cell surface nucleolin (NCL) interacts with RASSF2 via its RNA-binding domain, and NCL facilitates nuclear translocation of RASSF2. This nuclear transport of RASSF2 exacerbates endothelial cell pyroptosis and amplifies inflammatory responses in atherosclerosis.","method":"Immunoprecipitation-mass spectrometry (IP-MS), Co-immunoprecipitation, subcellular fractionation, NCL knockdown, in vivo atherosclerosis model","journal":"Atherosclerosis","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — IP-MS plus Co-IP validation and functional knockdown in vivo, single lab","pmids":["41895182"],"is_preprint":false},{"year":2026,"finding":"SETDB1 catalyzes H3K9me3 histone methylation at the RASSF2 promoter, transcriptionally repressing RASSF2 in ovarian cancer. RASSF2 stabilizes PTEN protein (demonstrated by Co-IP and cycloheximide chase assays) to inhibit PI3K/AKT pathway activation.","method":"Chromatin immunoprecipitation (ChIP) for SETDB1 and H3K9me3 at RASSF2 promoter, Co-immunoprecipitation (RASSF2-PTEN), cycloheximide chase assay, in vitro and in vivo overexpression/knockdown","journal":"Molecular genetics and genomics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP, Co-IP, and protein stability assay in single lab with multiple orthogonal methods","pmids":["42095894"],"is_preprint":false}],"current_model":"RASSF2 is a K-Ras-specific effector and tumor suppressor that binds K-Ras in a GTP-dependent manner and functions as a nuclear scaffolding protein; it activates proapoptotic MST1/MST2 kinases (protecting MST2 from proteolytic degradation), interacts with PAR-4 to facilitate its nuclear translocation and apoptotic function, suppresses NF-κB signaling by binding and inhibiting IKKα/β, is regulated by ERK2-mediated phosphorylation that controls its CRM-1-dependent nuclear export, associates with Rac GTPase regulators (including DOCK2) to sustain Rac activity, stabilizes PTEN to suppress PI3K/AKT signaling, and is frequently silenced by promoter CpG methylation (reinforced by SETDB1-driven H3K9me3) across multiple cancer types."},"narrative":{"mechanistic_narrative":"RASSF2 is a K-Ras-specific effector and tumor suppressor that links activated Ras to proapoptotic and growth-suppressive programs while shuttling between nucleus and cytoplasm [PMID:12732644, PMID:17891178]. It binds K-Ras directly in a GTP-dependent manner through its Ras-association domain, with this interaction required for its full proapoptotic activity, and loss of RASSF2 enhances K-Ras-driven transformation [PMID:12732644, PMID:18294275, PMID:16012945]. RASSF2 engages the proapoptotic Hippo kinases MST1 and MST2 via its SARAH domain, activating MST kinase activity and protecting MST2 from proteolytic degradation; reciprocally MST1 stabilizes RASSF2, and the MST-interaction domain is required for RASSF2-induced apoptosis [PMID:19525978, PMID:19962960, PMID:20920251]. Its localization is tightly controlled: a bipartite NLS drives nuclear accumulation needed for growth suppression, while ERK2-mediated phosphorylation licenses CRM-1-dependent nuclear export through a C-terminal NES, and import-defective RASSF2 fails to induce G1/S arrest or apoptosis [PMID:17891178, PMID:19555684]. In the nucleus and cytoplasm RASSF2 acts as a scaffold for additional apoptotic and signaling factors, forming a K-Ras-regulated complex with PAR-4 to promote PAR-4 nuclear translocation and apoptosis [PMID:20368356], and restraining oncogenic signaling by stabilizing PTEN to suppress PI3K/AKT and by binding IKKα/β to inhibit NF-κB during osteoclast and osteoblast differentiation [PMID:42095894, PMID:22227519]. Beyond canonical Hippo signaling, RASSF2 associates with Rac GTPase regulators including DOCK2 to sustain Rac activation and suppresses t(8;21) AML [PMID:32029705]. RASSF2 is recurrently silenced in cancer through promoter CpG-associated SETDB1-driven H3K9me3 and is targeted by miR-7 and the miR-200 family, integrating it into tumor-suppressive networks across multiple tissues [PMID:42095894, PMID:27901488, PMID:31565080].","teleology":[{"year":2003,"claim":"Established RASSF2 as a direct, GTP-dependent K-Ras effector, defining the molecular basis for its placement in Ras signaling rather than a generic Ras-association protein.","evidence":"Direct pulldown binding assays with GTP-dependence and cell-based apoptosis/cell cycle assays","pmids":["12732644"],"confidence":"High","gaps":["Did not resolve downstream effectors mediating apoptosis","Structural basis of K-Ras specificity over H-Ras not defined"]},{"year":2005,"claim":"Positioned RASSF2 functionally downstream of K-Ras as a negative regulator of transformation, showing its loss cooperates with oncogenic Ras.","evidence":"Colony formation, flow cytometry, and functional complementation in colorectal cancer cells","pmids":["16012945"],"confidence":"Medium","gaps":["Molecular mechanism of transformation suppression not defined","Single-lab cell-based evidence"]},{"year":2008,"claim":"Demonstrated that the Ras-association domain is required for RASSF2 proapoptotic function, tying its apoptotic role to Ras binding.","evidence":"Ras-association domain deletion mutagenesis with apoptosis assay in OSCC cells","pmids":["18294275"],"confidence":"Medium","gaps":["Does not identify the apoptotic effector engaged downstream of Ras binding","Single cell type"]},{"year":2007,"claim":"Identified a functional bipartite NLS and showed nuclear localization is required for tumor-suppressor activity, establishing RASSF2 as a localization-dependent suppressor.","evidence":"NLS mutagenesis with immunofluorescence and in vitro/in vivo growth suppression assays","pmids":["17891178"],"confidence":"High","gaps":["Nuclear targets of RASSF2 not identified","Mechanism coupling nuclear residence to growth suppression unresolved"]},{"year":2009,"claim":"Defined the RASSF2-MST1/2 axis: RASSF2 binds MST kinases via its SARAH domain, activates them, and protects MST2 from degradation, while MST1 reciprocally stabilizes RASSF2.","evidence":"Endogenous Co-IP, knockdown, kinase activity assays, and Mst1 knockout mice","pmids":["19525978","19962960"],"confidence":"High","gaps":["MST-independent JNK/apoptosis branch mechanism not fully resolved","Identity of the co-immunoprecipitating kinase phosphorylating RASSF2 not definitively established"]},{"year":2009,"claim":"Established that ERK2 phosphorylation controls CRM-1-dependent nuclear export through a defined C-terminal NES, coupling MAPK signaling to RASSF2 subcellular distribution and function.","evidence":"NES mutagenesis, leptomycin B, CRM-1 interaction, MAPK inhibition, in vitro phosphorylation, and cell cycle/apoptosis assays","pmids":["19555684"],"confidence":"High","gaps":["Phosphosite(s) on RASSF2 targeted by ERK2 not mapped","How nucleocytoplasmic shuttling integrates with MST/PAR-4 functions unclear"]},{"year":2010,"claim":"Showed RASSF2 scaffolds PAR-4 and drives its nuclear translocation in a K-Ras-regulated manner, providing a mechanism for PAR-4-dependent apoptosis.","evidence":"Endogenous Co-IP, subcellular fractionation, and apoptosis assays in prostate tumor cells","pmids":["20368356"],"confidence":"High","gaps":["How K-Ras regulation switches the RASSF2-PAR-4 interaction is not defined","Relationship to MST-dependent apoptosis not integrated"]},{"year":2010,"claim":"Confirmed the MST-interaction domain is functionally required for RASSF2-induced apoptosis in a distinct cancer type, generalizing the MST-dependence of its apoptotic activity.","evidence":"MST-interaction domain deletion with apoptosis assay in thyroid cancer cells","pmids":["20920251"],"confidence":"Medium","gaps":["Does not separate MST binding from MST activation","Single-lab evidence"]},{"year":2012,"claim":"Linked RASSF2 loss to AKT activation and validated endogenous RASSF2-K-Ras complexes in lung cancer, connecting RASSF2 to PI3K/AKT signaling.","evidence":"RNAi knockdown, endogenous Co-IP, AKT phosphorylation, and growth/invasion assays","pmids":["22693671"],"confidence":"Medium","gaps":["Mechanism linking RASSF2 to AKT not established at this stage","Single-lab readouts"]},{"year":2012,"claim":"Revealed an in vivo physiological role in bone remodeling and hematopoiesis and a mechanism whereby RASSF2 binds and inhibits IKKα/β to restrain NF-κB during differentiation.","evidence":"Rassf2 knockout mice, in vitro differentiation, RASSF2-IKK Co-IP, IKK kinase assay, and dominant-negative rescue","pmids":["22227519"],"confidence":"High","gaps":["How NF-κB suppression relates to RASSF2 apoptotic/Ras functions unclear","Structural basis of IKK inhibition not defined"]},{"year":2016,"claim":"Expanded the RASSF2 interactome to C1QBP and Vimentin in a K-Ras-modulated manner, linking RASSF2/K-Ras to vimentin acetylation.","evidence":"Proteomics/MS pulldown with Co-IP validation and acetylation assay","pmids":["26999212"],"confidence":"Medium","gaps":["Functional consequence of C1QBP and Vimentin binding not established","Mechanism of vimentin acetylation control unresolved"]},{"year":2017,"claim":"Placed RASSF2 in a miR-7-RASSF2-PAR-4 axis in cancer-associated fibroblasts, showing its repression alters the tumor microenvironment.","evidence":"miRNA gain/loss, co-culture, target validation, and proliferation/migration assays","pmids":["27901488"],"confidence":"Medium","gaps":["Direct mechanism of PAR-4 secretion control not defined","Single-lab co-culture model"]},{"year":2019,"claim":"Identified the miR-200 family as a direct regulator of RASSF2 mRNA coupling its levels to ERK/MAPK signaling, adding a feedback layer to RASSF2 regulation.","evidence":"miRNA gain/loss, knockdown, and ERK phosphorylation/proliferation readouts in colon cancer cells","pmids":["31565080"],"confidence":"Medium","gaps":["Direct biochemical target binding not structurally validated","Single cell line"]},{"year":2020,"claim":"Showed RASSF2 suppresses t(8;21) AML through MST1/2 interaction independent of canonical Hippo signaling and uncovered association with Rac regulators including DOCK2 to sustain Rac activation.","evidence":"AML re-expression models in vitro/in vivo, BioID proximity labeling, Co-IP, and Rac activation assays","pmids":["32029705"],"confidence":"High","gaps":["Direct vs indirect nature of RASSF2-DOCK2 association not resolved","How RASSF2 promotes Rac activation mechanistically unclear"]},{"year":2023,"claim":"Extended RASSF2 function to oxidative stress-induced apoptosis in lens epithelium via AKT Ser473 regulation, showing context-dependent proapoptotic signaling.","evidence":"Overexpression/knockdown, AKT phosphorylation western blots, and mouse model","pmids":["37979829"],"confidence":"Medium","gaps":["Direction of AKT regulation appears context-specific and is not mechanistically reconciled with tumor-suppressor PTEN/AKT findings","Single-lab study"]},{"year":2026,"claim":"Defined a PTEN-stabilizing mechanism for RASSF2-mediated PI3K/AKT suppression and identified SETDB1-driven H3K9me3 as a transcriptional silencing mechanism in ovarian cancer.","evidence":"ChIP for SETDB1/H3K9me3 at the RASSF2 promoter, RASSF2-PTEN Co-IP, and cycloheximide chase assays","pmids":["42095894"],"confidence":"Medium","gaps":["Mechanism by which RASSF2 stabilizes PTEN not defined","Single-lab evidence"]},{"year":2026,"claim":"Showed cell-surface nucleolin binds RASSF2 and drives its nuclear translocation to promote endothelial pyroptosis, implicating RASSF2 in vascular inflammation.","evidence":"IP-MS, Co-IP, subcellular fractionation, NCL knockdown, and in vivo atherosclerosis model","pmids":["41895182"],"confidence":"Medium","gaps":["How nuclear RASSF2 triggers pyroptosis mechanistically is unresolved","Single-lab study"]},{"year":null,"claim":"It remains unresolved how RASSF2 integrates its distinct effector branches (MST/Hippo, PAR-4, IKK/NF-κB, PTEN/AKT, Rac/DOCK2) into a unified, context-dependent program, and whether its apparently opposing roles in apoptosis suppression versus promotion reflect tissue-specific partner availability.","evidence":"","pmids":[],"confidence":"Low","gaps":["No structural model of RASSF2 with its multiple partners","No unified framework reconciling tumor-suppressive and proinflammatory/proapoptotic roles across tissues"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[3,6,17]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[4,10,3]}],"localization":[{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[2,3,5]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[3,5]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,9,13]},{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[0,3,6]}],"complexes":[],"partners":["KRAS","MST1","MST2","PAWR","IKBKB","DOCK2","PTEN","NCL"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"P50749","full_name":"Ras association domain-containing protein 2","aliases":[],"length_aa":326,"mass_kda":37.8,"function":"Potential tumor suppressor. Acts as a KRAS-specific effector protein. May promote apoptosis and cell cycle arrest. Stabilizes STK3/MST2 by protecting it from proteasomal degradation","subcellular_location":"Nucleus; Cytoplasm; Chromosome, centromere, kinetochore","url":"https://www.uniprot.org/uniprotkb/P50749/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/RASSF2","classification":"Not Classified","n_dependent_lines":1,"n_total_lines":1208,"dependency_fraction":0.0008278145695364238},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"STK4","stoichiometry":0.2},{"gene":"TAOK1","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/RASSF2","total_profiled":1310},"omim":[{"mim_id":"612620","title":"RAS ASSOCIATION DOMAIN FAMILY, MEMBER 6; RASSF6","url":"https://www.omim.org/entry/612620"},{"mim_id":"610559","title":"RAS ASSOCIATION DOMAIN FAMILY, MEMBER 4; RASSF4","url":"https://www.omim.org/entry/610559"},{"mim_id":"609492","title":"RAS ASSOCIATION DOMAIN FAMILY PROTEIN 2; RASSF2","url":"https://www.omim.org/entry/609492"},{"mim_id":"601936","title":"PRKC, APOPTOSIS, WT1, REGULATOR; PAWR","url":"https://www.omim.org/entry/601936"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Nucleoplasm","reliability":"Supported"},{"location":"Golgi apparatus","reliability":"Additional"},{"location":"Plasma membrane","reliability":"Additional"},{"location":"Cytosol","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"brain","ntpm":286.3},{"tissue":"retina","ntpm":150.9}],"url":"https://www.proteinatlas.org/search/RASSF2"},"hgnc":{"alias_symbol":["KIAA0168","CENP-34"],"prev_symbol":[]},"alphafold":{"accession":"P50749","domains":[{"cath_id":"3.10.20.90","chopping":"10-68_184-269","consensus_level":"high","plddt":95.3597,"start":10,"end":269},{"cath_id":"1.20.5","chopping":"279-326","consensus_level":"medium","plddt":92.6652,"start":279,"end":326}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P50749","model_url":"https://alphafold.ebi.ac.uk/files/AF-P50749-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P50749-F1-predicted_aligned_error_v6.png","plddt_mean":77.88},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=RASSF2","jax_strain_url":"https://www.jax.org/strain/search?query=RASSF2"},"sequence":{"accession":"P50749","fasta_url":"https://rest.uniprot.org/uniprotkb/P50749.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P50749/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P50749"}},"corpus_meta":[{"pmid":"12732644","id":"PMC_12732644","title":"RASSF2 is a novel K-Ras-specific effector and potential tumor suppressor.","date":"2003","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/12732644","citation_count":151,"is_preprint":false},{"pmid":"16012945","id":"PMC_16012945","title":"The Ras effector RASSF2 is a novel tumor-suppressor gene in human colorectal cancer.","date":"2005","source":"Gastroenterology","url":"https://pubmed.ncbi.nlm.nih.gov/16012945","citation_count":114,"is_preprint":false},{"pmid":"19525978","id":"PMC_19525978","title":"RASSF2 associates with and stabilizes the proapoptotic kinase MST2.","date":"2009","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/19525978","citation_count":70,"is_preprint":false},{"pmid":"16265349","id":"PMC_16265349","title":"RASSF2, a potential tumour suppressor, is silenced by CpG island hypermethylation in gastric cancer.","date":"2005","source":"British journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/16265349","citation_count":62,"is_preprint":false},{"pmid":"20920251","id":"PMC_20920251","title":"Frequent epigenetic inactivation of RASSF2 in thyroid cancer and functional consequences.","date":"2010","source":"Molecular cancer","url":"https://pubmed.ncbi.nlm.nih.gov/20920251","citation_count":49,"is_preprint":false},{"pmid":"19962960","id":"PMC_19962960","title":"Role of the tumor suppressor RASSF2 in regulation of MST1 kinase activity.","date":"2009","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/19962960","citation_count":47,"is_preprint":false},{"pmid":"27901488","id":"PMC_27901488","title":"Cancer-associated fibroblasts promote cancer cell growth through a miR-7-RASSF2-PAR-4 axis in the tumor microenvironment.","date":"2017","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/27901488","citation_count":47,"is_preprint":false},{"pmid":"31565080","id":"PMC_31565080","title":"The microRNA-200 family acts as an oncogene in colorectal cancer by inhibiting the tumor suppressor RASSF2.","date":"2019","source":"Oncology letters","url":"https://pubmed.ncbi.nlm.nih.gov/31565080","citation_count":44,"is_preprint":false},{"pmid":"17891178","id":"PMC_17891178","title":"Epigenetic regulation of the ras effector/tumour suppressor RASSF2 in breast and lung cancer.","date":"2007","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/17891178","citation_count":43,"is_preprint":false},{"pmid":"18294275","id":"PMC_18294275","title":"Epigenetic inactivation of RASSF2 in oral squamous cell carcinoma.","date":"2008","source":"Cancer science","url":"https://pubmed.ncbi.nlm.nih.gov/18294275","citation_count":42,"is_preprint":false},{"pmid":"22227519","id":"PMC_22227519","title":"Ablation of Rassf2 induces bone defects and subsequent haematopoietic anomalies in mice.","date":"2012","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/22227519","citation_count":36,"is_preprint":false},{"pmid":"23542458","id":"PMC_23542458","title":"RASSF2 hypermethylation is present and related to shorter survival in squamous cervical cancer.","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/23542458","citation_count":29,"is_preprint":false},{"pmid":"17549418","id":"PMC_17549418","title":"Epigenetic inactivation of the RAS-effector gene RASSF2 in lung cancers.","date":"2007","source":"International journal of oncology","url":"https://pubmed.ncbi.nlm.nih.gov/17549418","citation_count":27,"is_preprint":false},{"pmid":"20368356","id":"PMC_20368356","title":"The Ras effector RASSF2 controls the PAR-4 tumor suppressor.","date":"2010","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/20368356","citation_count":27,"is_preprint":false},{"pmid":"26782783","id":"PMC_26782783","title":"Comparative Modeling, Molecular Docking, and Revealing of Potential Binding Pockets of RASSF2; a Candidate Cancer Gene.","date":"2016","source":"Interdisciplinary sciences, computational life sciences","url":"https://pubmed.ncbi.nlm.nih.gov/26782783","citation_count":20,"is_preprint":false},{"pmid":"22693671","id":"PMC_22693671","title":"Loss of RASSF2 Enhances Tumorigencity of Lung Cancer Cells and Confers Resistance to Chemotherapy.","date":"2012","source":"Molecular biology 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Partners.","date":"2016","source":"Cancers","url":"https://pubmed.ncbi.nlm.nih.gov/26999212","citation_count":10,"is_preprint":false},{"pmid":"37979829","id":"PMC_37979829","title":"Epigallocatechin gallate delays age-related cataract development via the RASSF2/AKT pathway.","date":"2023","source":"European journal of pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/37979829","citation_count":8,"is_preprint":false},{"pmid":"24519048","id":"PMC_24519048","title":"[Detection of RASSF2 and sFRP1 promoter region methylation in sporadic colorectal cancer patients].","date":"2014","source":"Zhonghua wei chang wai ke za zhi = Chinese journal of gastrointestinal surgery","url":"https://pubmed.ncbi.nlm.nih.gov/24519048","citation_count":4,"is_preprint":false},{"pmid":"31453724","id":"PMC_31453724","title":"Association Between RASSF2 Methylation and Gastric Cancer: A PRISMA-Compliant Systematic Review and Meta-Analysis.","date":"2019","source":"DNA and cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/31453724","citation_count":2,"is_preprint":false},{"pmid":"38036076","id":"PMC_38036076","title":"Hyperhomocysteinemia may aggravate abdominal aortic aneurysm formation by up-regulating RASSF2.","date":"2023","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/38036076","citation_count":2,"is_preprint":false},{"pmid":"41895182","id":"PMC_41895182","title":"Cell surface nucleolin promotes endothelial cell pyroptosis in atherosclerosis through RASSF2.","date":"2026","source":"Atherosclerosis","url":"https://pubmed.ncbi.nlm.nih.gov/41895182","citation_count":0,"is_preprint":false},{"pmid":"42020562","id":"PMC_42020562","title":"RASSF2 promoter hypermethylation determines malignant and microenvironmental features in lung cancer.","date":"2026","source":"Genes and immunity","url":"https://pubmed.ncbi.nlm.nih.gov/42020562","citation_count":0,"is_preprint":false},{"pmid":"42095894","id":"PMC_42095894","title":"SETDB1 induces H3K9me3 methylation modification of RASSF2 to inhibit PTEN and facilitate ovarian cancer progression.","date":"2026","source":"Molecular genetics and genomics : MGG","url":"https://pubmed.ncbi.nlm.nih.gov/42095894","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":15624,"output_tokens":4581,"usd":0.057793,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":12512,"output_tokens":5030,"usd":0.094155,"stage2_stop_reason":"end_turn"},"total_usd":0.151948,"stage1_batch_id":"msgbatch_01CNoDbkChHEkgrJnL7BwtGJ","stage2_batch_id":"msgbatch_016Y4x9E9Wg5VQDANj9quSQD","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2003,\n      \"finding\": \"RASSF2 binds directly to K-Ras in a GTP-dependent manner via the Ras effector domain, but only weakly interacts with H-Ras, establishing it as a K-Ras-specific effector. RASSF2 promotes apoptosis and cell cycle arrest.\",\n      \"method\": \"Direct binding assay (pulldown), GTP-dependence assay, cell-based apoptosis and cell cycle assays\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal binding assay with GTP-dependence, replicated in subsequent studies across multiple labs\",\n      \"pmids\": [\"12732644\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"RASSF2 induces morphological changes and apoptosis when ectopically expressed in colorectal cancer cells, and inactivation of RASSF2 enhances K-Ras-induced oncogenic transformation, placing RASSF2 downstream of K-Ras as a negative regulator of transformation.\",\n      \"method\": \"Colony formation assay, flow cytometry, immunofluorescence microscopy, functional complementation\",\n      \"journal\": \"Gastroenterology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal cell-based assays in a single lab\",\n      \"pmids\": [\"16012945\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"RASSF2 contains a functional bipartite nuclear localization signal (NLS); mutation of the NLS abolishes nuclear localization and significantly diminishes its tumor suppressor (growth suppression) activity, demonstrating that nuclear localization is required for full RASSF2 function.\",\n      \"method\": \"Site-directed mutagenesis of NLS, immunofluorescence localization, in vitro and in vivo growth suppression assays\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — mutagenesis combined with localization and functional growth suppression assays in vitro and in vivo\",\n      \"pmids\": [\"17891178\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"RASSF2 associates with the proapoptotic kinases MST1 and MST2 via its SARAH domain, and this interaction is confirmed at endogenous levels. RASSF2 co-immunoprecipitates active MST1/2, is phosphorylated by a co-immunoprecipitating kinase (likely MST1/2), and stabilizes MST2 by protecting it from proteolytic degradation. RASSF2 alone localizes to the nucleus, but co-expression with MST1 or MST2 results in cytoplasmic co-localization.\",\n      \"method\": \"Co-immunoprecipitation (endogenous), immunofluorescence co-localization, stable/transient expression, RASSF2 knockdown, kinase activity assay\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — endogenous Co-IP, multiple orthogonal methods, knockdown and overexpression, replicated in subsequent studies\",\n      \"pmids\": [\"19525978\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"MST1 regulates RASSF2 protein stability: knockdown of MST1 markedly destabilizes RASSF2, and Mst1-deficient mice show reduced Rassf2 protein levels. Conversely, RASSF2 activates MST1 kinase activity through formation of a RASSF2-MST1 complex. RASSF2 also activates JNK signaling and induces apoptosis in an MST1-independent manner.\",\n      \"method\": \"siRNA knockdown, Mst1 knockout mice, kinase activity assay, complex formation (Co-IP), apoptosis assay, JNK pathway assay\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic (knockout mice) plus biochemical (kinase assay, Co-IP) evidence, multiple orthogonal methods\",\n      \"pmids\": [\"19962960\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"RASSF2 shuttles between nucleus and cytoplasm via a CRM-1-dependent nuclear export mechanism. A functional nuclear export signal (NES) resides in amino acids 240–260 (C-terminus); mutation of conserved residues Ile254, Val257, and Leu259 impairs nuclear export and interaction with CRM-1. ERK2 (MAPK) phosphorylates RASSF2 and this phosphorylation is required for nuclear export; inhibition of MAPK pathway blocks RASSF2 export. Nuclear import-defective RASSF2 fails to induce G1/S cell cycle arrest or apoptosis.\",\n      \"method\": \"NES mutagenesis, leptomycin B treatment, CRM-1 interaction assay, MAPK inhibitor treatment, in vitro phosphorylation assay, cell cycle and apoptosis assays\",\n      \"journal\": \"Experimental cell research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — mutagenesis, pharmacological inhibition, in vitro phosphorylation, and functional phenotype in one study\",\n      \"pmids\": [\"19555684\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"RASSF2 forms a direct and endogenous complex with PAR-4 (prostate apoptosis response protein 4). This interaction is regulated by K-Ras and is essential for the full apoptotic effects of PAR-4. RASSF2 (a primarily nuclear protein) modulates nuclear translocation of PAR-4 from cytoplasm to nucleus in prostate tumor cells, providing a mechanism for PAR-4's biological apoptotic effects.\",\n      \"method\": \"Co-immunoprecipitation (endogenous), subcellular fractionation/localization, functional apoptosis assay\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — endogenous Co-IP, localization experiments, and functional apoptosis assay in one rigorous study\",\n      \"pmids\": [\"20368356\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Deletion of the MST interaction domain of RASSF2 significantly reduces RASSF2-induced apoptosis in thyroid cancer cells, establishing that the MST interaction domain is functionally required for RASSF2-mediated apoptosis.\",\n      \"method\": \"Domain deletion mutagenesis, apoptosis assay in thyroid cancer cells\",\n      \"journal\": \"Molecular cancer\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — deletion mutagenesis with quantitative apoptosis readout in a single lab\",\n      \"pmids\": [\"20920251\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"A RASSF2 deletion mutant lacking the Ras-association domain, unable to interact with Ras, exhibits less pro-apoptotic activity than the full-length protein, demonstrating that the pro-apoptotic activity of RASSF2 is related to its association with Ras.\",\n      \"method\": \"Deletion mutagenesis, apoptosis assay in OSCC cells\",\n      \"journal\": \"Cancer science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — domain deletion mutagenesis combined with functional apoptosis assay, single lab\",\n      \"pmids\": [\"18294275\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Loss of RASSF2 expression in lung cancer cells correlates with increased levels of activated AKT, indicating RASSF2 modulates Ras/AKT signaling. RASSF2 and K-Ras form an endogenous complex (confirmed by Co-IP), validating RASSF2 as a bona fide K-Ras effector in lung cancer cells.\",\n      \"method\": \"RNAi knockdown, Co-immunoprecipitation (endogenous), AKT phosphorylation assay, cell proliferation/invasion/anchorage-independent growth assays\",\n      \"journal\": \"Molecular biology international\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — endogenous Co-IP and signaling assay, single lab with multiple functional readouts\",\n      \"pmids\": [\"22693671\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Rassf2 knockout mice develop bone remodeling defects and haematopoietic anomalies. RASSF2 ablation suppresses osteoblastogenesis and promotes osteoclastogenesis in vitro. RASSF2 associates with IKKα and IKKβ and suppresses IKK kinase activity, thereby restraining NF-κB signaling during osteoclast and osteoblast differentiation. Reintroduction of RASSF2 or dominant-negative IKK normalizes differentiation in Rassf2-/- precursors.\",\n      \"method\": \"Knockout mouse model, bone marrow transplantation, in vitro differentiation assay, Co-immunoprecipitation (RASSF2-IKKα/β), IKK kinase activity assay, dominant-negative rescue\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — genetic knockout model, Co-IP, kinase assay, rescue experiment, multiple orthogonal methods in one rigorous study\",\n      \"pmids\": [\"22227519\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Proteomics-based screening identified novel RASSF2 interaction partners including C1QBP and Vimentin. The RASSF2-C1QBP interaction is enhanced by K-Ras, while the RASSF2-Vimentin interaction is reduced by K-Ras. RASSF2/K-Ras regulates the acetylation of Vimentin.\",\n      \"method\": \"Proteomics/MS pulldown, Co-immunoprecipitation validation, acetylation assay\",\n      \"journal\": \"Cancers\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — proteomics screen validated by Co-IP for two interactions and acetylation assay, single lab\",\n      \"pmids\": [\"26999212\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"RASSF2 is a direct target of miR-7 in cancer-associated fibroblasts. Overexpression of miR-7 down-regulates RASSF2, which reduces PAR-4 secretion from fibroblasts and enhances proliferation and migration of co-cultured cancer cells, placing RASSF2 in a miR-7-RASSF2-PAR-4 signaling axis.\",\n      \"method\": \"miRNA overexpression/inhibition, co-culture assay, bioinformatic target validation, functional proliferation/migration assay\",\n      \"journal\": \"Oncotarget\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — functional miRNA/target assays with co-culture readout, single lab\",\n      \"pmids\": [\"27901488\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"RASSF2 suppresses t(8;21) AML development, and its function depends on interaction with Hippo kinases MST1 and MST2, but is independent of canonical Hippo pathway signaling. Proximity-based biotin labeling revealed RASSF2 associates with Rac GTPase-related proteins including the guanine nucleotide exchange factor DOCK2. RASSF2 knockdown impairs Rac GTPase activation.\",\n      \"method\": \"Re-expression in AML models (in vitro and in vivo), proximity-based biotin labeling (BioID), Co-immunoprecipitation, Rac GTPase activation assay, MST1/2 interaction studies\",\n      \"journal\": \"Blood cancer journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — BioID proteomics, Co-IP, functional rescue in multiple AML models, Rac activation assay, multiple orthogonal methods\",\n      \"pmids\": [\"32029705\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"The miR-200 family directly targets RASSF2 mRNA; knockdown of the miR-200 family in HT-29 colon cancer cells increases RASSF2 expression and decreases ERK phosphorylation (MAPK/ERK signaling), while increased miR-200 expression decreases RASSF2 and increases ERK phosphorylation.\",\n      \"method\": \"siRNA knockdown, miRNA gain/loss of function, western blot for ERK phosphorylation, cell proliferation assay\",\n      \"journal\": \"Oncology letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — direct miRNA-target functional experiments with signaling readout, single lab\",\n      \"pmids\": [\"31565080\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"RASSF2 mediates oxidative stress-induced apoptosis in lens epithelial cells by regulating AKT (Ser473) phosphorylation. Overexpression of RASSF2 promotes apoptosis under oxidative conditions, while knockdown reduces it. EGCG inhibits this pathway by reducing RASSF2 levels, thereby suppressing AKT phosphorylation and apoptosis.\",\n      \"method\": \"Overexpression and siRNA knockdown in lens epithelial cells, western blot for AKT Ser473 phosphorylation, mouse model\",\n      \"journal\": \"European journal of pharmacology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — overexpression/knockdown with phosphorylation readout, in vitro and in vivo, single lab\",\n      \"pmids\": [\"37979829\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"Cell surface nucleolin (NCL) interacts with RASSF2 via its RNA-binding domain, and NCL facilitates nuclear translocation of RASSF2. This nuclear transport of RASSF2 exacerbates endothelial cell pyroptosis and amplifies inflammatory responses in atherosclerosis.\",\n      \"method\": \"Immunoprecipitation-mass spectrometry (IP-MS), Co-immunoprecipitation, subcellular fractionation, NCL knockdown, in vivo atherosclerosis model\",\n      \"journal\": \"Atherosclerosis\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — IP-MS plus Co-IP validation and functional knockdown in vivo, single lab\",\n      \"pmids\": [\"41895182\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"SETDB1 catalyzes H3K9me3 histone methylation at the RASSF2 promoter, transcriptionally repressing RASSF2 in ovarian cancer. RASSF2 stabilizes PTEN protein (demonstrated by Co-IP and cycloheximide chase assays) to inhibit PI3K/AKT pathway activation.\",\n      \"method\": \"Chromatin immunoprecipitation (ChIP) for SETDB1 and H3K9me3 at RASSF2 promoter, Co-immunoprecipitation (RASSF2-PTEN), cycloheximide chase assay, in vitro and in vivo overexpression/knockdown\",\n      \"journal\": \"Molecular genetics and genomics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP, Co-IP, and protein stability assay in single lab with multiple orthogonal methods\",\n      \"pmids\": [\"42095894\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"RASSF2 is a K-Ras-specific effector and tumor suppressor that binds K-Ras in a GTP-dependent manner and functions as a nuclear scaffolding protein; it activates proapoptotic MST1/MST2 kinases (protecting MST2 from proteolytic degradation), interacts with PAR-4 to facilitate its nuclear translocation and apoptotic function, suppresses NF-κB signaling by binding and inhibiting IKKα/β, is regulated by ERK2-mediated phosphorylation that controls its CRM-1-dependent nuclear export, associates with Rac GTPase regulators (including DOCK2) to sustain Rac activity, stabilizes PTEN to suppress PI3K/AKT signaling, and is frequently silenced by promoter CpG methylation (reinforced by SETDB1-driven H3K9me3) across multiple cancer types.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"RASSF2 is a K-Ras-specific effector and tumor suppressor that links activated Ras to proapoptotic and growth-suppressive programs while shuttling between nucleus and cytoplasm [#0, #2]. It binds K-Ras directly in a GTP-dependent manner through its Ras-association domain, with this interaction required for its full proapoptotic activity, and loss of RASSF2 enhances K-Ras-driven transformation [#0, #8, #1]. RASSF2 engages the proapoptotic Hippo kinases MST1 and MST2 via its SARAH domain, activating MST kinase activity and protecting MST2 from proteolytic degradation; reciprocally MST1 stabilizes RASSF2, and the MST-interaction domain is required for RASSF2-induced apoptosis [#3, #4, #7]. Its localization is tightly controlled: a bipartite NLS drives nuclear accumulation needed for growth suppression, while ERK2-mediated phosphorylation licenses CRM-1-dependent nuclear export through a C-terminal NES, and import-defective RASSF2 fails to induce G1/S arrest or apoptosis [#2, #5]. In the nucleus and cytoplasm RASSF2 acts as a scaffold for additional apoptotic and signaling factors, forming a K-Ras-regulated complex with PAR-4 to promote PAR-4 nuclear translocation and apoptosis [#6], and restraining oncogenic signaling by stabilizing PTEN to suppress PI3K/AKT and by binding IKK\\u03b1/\\u03b2 to inhibit NF-\\u03baB during osteoclast and osteoblast differentiation [#17, #10]. Beyond canonical Hippo signaling, RASSF2 associates with Rac GTPase regulators including DOCK2 to sustain Rac activation and suppresses t(8;21) AML [#13]. RASSF2 is recurrently silenced in cancer through promoter CpG-associated SETDB1-driven H3K9me3 and is targeted by miR-7 and the miR-200 family, integrating it into tumor-suppressive networks across multiple tissues [#17, #12, #14].\",\n  \"teleology\": [\n    {\n      \"year\": 2003,\n      \"claim\": \"Established RASSF2 as a direct, GTP-dependent K-Ras effector, defining the molecular basis for its placement in Ras signaling rather than a generic Ras-association protein.\",\n      \"evidence\": \"Direct pulldown binding assays with GTP-dependence and cell-based apoptosis/cell cycle assays\",\n      \"pmids\": [\"12732644\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not resolve downstream effectors mediating apoptosis\", \"Structural basis of K-Ras specificity over H-Ras not defined\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Positioned RASSF2 functionally downstream of K-Ras as a negative regulator of transformation, showing its loss cooperates with oncogenic Ras.\",\n      \"evidence\": \"Colony formation, flow cytometry, and functional complementation in colorectal cancer cells\",\n      \"pmids\": [\"16012945\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular mechanism of transformation suppression not defined\", \"Single-lab cell-based evidence\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Demonstrated that the Ras-association domain is required for RASSF2 proapoptotic function, tying its apoptotic role to Ras binding.\",\n      \"evidence\": \"Ras-association domain deletion mutagenesis with apoptosis assay in OSCC cells\",\n      \"pmids\": [\"18294275\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Does not identify the apoptotic effector engaged downstream of Ras binding\", \"Single cell type\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Identified a functional bipartite NLS and showed nuclear localization is required for tumor-suppressor activity, establishing RASSF2 as a localization-dependent suppressor.\",\n      \"evidence\": \"NLS mutagenesis with immunofluorescence and in vitro/in vivo growth suppression assays\",\n      \"pmids\": [\"17891178\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Nuclear targets of RASSF2 not identified\", \"Mechanism coupling nuclear residence to growth suppression unresolved\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Defined the RASSF2-MST1/2 axis: RASSF2 binds MST kinases via its SARAH domain, activates them, and protects MST2 from degradation, while MST1 reciprocally stabilizes RASSF2.\",\n      \"evidence\": \"Endogenous Co-IP, knockdown, kinase activity assays, and Mst1 knockout mice\",\n      \"pmids\": [\"19525978\", \"19962960\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"MST-independent JNK/apoptosis branch mechanism not fully resolved\", \"Identity of the co-immunoprecipitating kinase phosphorylating RASSF2 not definitively established\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Established that ERK2 phosphorylation controls CRM-1-dependent nuclear export through a defined C-terminal NES, coupling MAPK signaling to RASSF2 subcellular distribution and function.\",\n      \"evidence\": \"NES mutagenesis, leptomycin B, CRM-1 interaction, MAPK inhibition, in vitro phosphorylation, and cell cycle/apoptosis assays\",\n      \"pmids\": [\"19555684\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Phosphosite(s) on RASSF2 targeted by ERK2 not mapped\", \"How nucleocytoplasmic shuttling integrates with MST/PAR-4 functions unclear\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Showed RASSF2 scaffolds PAR-4 and drives its nuclear translocation in a K-Ras-regulated manner, providing a mechanism for PAR-4-dependent apoptosis.\",\n      \"evidence\": \"Endogenous Co-IP, subcellular fractionation, and apoptosis assays in prostate tumor cells\",\n      \"pmids\": [\"20368356\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How K-Ras regulation switches the RASSF2-PAR-4 interaction is not defined\", \"Relationship to MST-dependent apoptosis not integrated\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Confirmed the MST-interaction domain is functionally required for RASSF2-induced apoptosis in a distinct cancer type, generalizing the MST-dependence of its apoptotic activity.\",\n      \"evidence\": \"MST-interaction domain deletion with apoptosis assay in thyroid cancer cells\",\n      \"pmids\": [\"20920251\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Does not separate MST binding from MST activation\", \"Single-lab evidence\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Linked RASSF2 loss to AKT activation and validated endogenous RASSF2-K-Ras complexes in lung cancer, connecting RASSF2 to PI3K/AKT signaling.\",\n      \"evidence\": \"RNAi knockdown, endogenous Co-IP, AKT phosphorylation, and growth/invasion assays\",\n      \"pmids\": [\"22693671\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism linking RASSF2 to AKT not established at this stage\", \"Single-lab readouts\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Revealed an in vivo physiological role in bone remodeling and hematopoiesis and a mechanism whereby RASSF2 binds and inhibits IKK\\u03b1/\\u03b2 to restrain NF-\\u03baB during differentiation.\",\n      \"evidence\": \"Rassf2 knockout mice, in vitro differentiation, RASSF2-IKK Co-IP, IKK kinase assay, and dominant-negative rescue\",\n      \"pmids\": [\"22227519\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How NF-\\u03baB suppression relates to RASSF2 apoptotic/Ras functions unclear\", \"Structural basis of IKK inhibition not defined\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Expanded the RASSF2 interactome to C1QBP and Vimentin in a K-Ras-modulated manner, linking RASSF2/K-Ras to vimentin acetylation.\",\n      \"evidence\": \"Proteomics/MS pulldown with Co-IP validation and acetylation assay\",\n      \"pmids\": [\"26999212\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional consequence of C1QBP and Vimentin binding not established\", \"Mechanism of vimentin acetylation control unresolved\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Placed RASSF2 in a miR-7-RASSF2-PAR-4 axis in cancer-associated fibroblasts, showing its repression alters the tumor microenvironment.\",\n      \"evidence\": \"miRNA gain/loss, co-culture, target validation, and proliferation/migration assays\",\n      \"pmids\": [\"27901488\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct mechanism of PAR-4 secretion control not defined\", \"Single-lab co-culture model\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Identified the miR-200 family as a direct regulator of RASSF2 mRNA coupling its levels to ERK/MAPK signaling, adding a feedback layer to RASSF2 regulation.\",\n      \"evidence\": \"miRNA gain/loss, knockdown, and ERK phosphorylation/proliferation readouts in colon cancer cells\",\n      \"pmids\": [\"31565080\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct biochemical target binding not structurally validated\", \"Single cell line\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Showed RASSF2 suppresses t(8;21) AML through MST1/2 interaction independent of canonical Hippo signaling and uncovered association with Rac regulators including DOCK2 to sustain Rac activation.\",\n      \"evidence\": \"AML re-expression models in vitro/in vivo, BioID proximity labeling, Co-IP, and Rac activation assays\",\n      \"pmids\": [\"32029705\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct vs indirect nature of RASSF2-DOCK2 association not resolved\", \"How RASSF2 promotes Rac activation mechanistically unclear\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Extended RASSF2 function to oxidative stress-induced apoptosis in lens epithelium via AKT Ser473 regulation, showing context-dependent proapoptotic signaling.\",\n      \"evidence\": \"Overexpression/knockdown, AKT phosphorylation western blots, and mouse model\",\n      \"pmids\": [\"37979829\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direction of AKT regulation appears context-specific and is not mechanistically reconciled with tumor-suppressor PTEN/AKT findings\", \"Single-lab study\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Defined a PTEN-stabilizing mechanism for RASSF2-mediated PI3K/AKT suppression and identified SETDB1-driven H3K9me3 as a transcriptional silencing mechanism in ovarian cancer.\",\n      \"evidence\": \"ChIP for SETDB1/H3K9me3 at the RASSF2 promoter, RASSF2-PTEN Co-IP, and cycloheximide chase assays\",\n      \"pmids\": [\"42095894\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism by which RASSF2 stabilizes PTEN not defined\", \"Single-lab evidence\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Showed cell-surface nucleolin binds RASSF2 and drives its nuclear translocation to promote endothelial pyroptosis, implicating RASSF2 in vascular inflammation.\",\n      \"evidence\": \"IP-MS, Co-IP, subcellular fractionation, NCL knockdown, and in vivo atherosclerosis model\",\n      \"pmids\": [\"41895182\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"How nuclear RASSF2 triggers pyroptosis mechanistically is unresolved\", \"Single-lab study\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"It remains unresolved how RASSF2 integrates its distinct effector branches (MST/Hippo, PAR-4, IKK/NF-\\u03baB, PTEN/AKT, Rac/DOCK2) into a unified, context-dependent program, and whether its apparently opposing roles in apoptosis suppression versus promotion reflect tissue-specific partner availability.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No structural model of RASSF2 with its multiple partners\", \"No unified framework reconciling tumor-suppressive and proinflammatory/proapoptotic roles across tissues\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [3, 6, 17]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [4, 10, 3]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [2, 3, 5]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [3, 5]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 9, 13]},\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [0, 3, 6]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"KRAS\", \"MST1\", \"MST2\", \"PAWR\", \"IKBKB\", \"DOCK2\", \"PTEN\", \"NCL\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"tie","faith_supported":7,"faith_total":7,"faith_pct":100.0}}