{"gene":"RASSF2","run_date":"2026-04-28T19:45:45","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, identifying it as a K-Ras-specific effector that promotes apoptosis and cell cycle arrest.","method":"Direct binding assay (GTP-dependent interaction), overexpression with apoptosis/cell cycle readouts","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 — direct binding assay with GTP-dependence specificity, replicated by multiple subsequent studies","pmids":["12732644"],"is_preprint":false},{"year":2005,"finding":"RASSF2 expression induces morphological changes and apoptosis in colorectal cancer cells; a deletion mutant lacking the RA domain shows reduced pro-apoptotic activity, indicating the Ras interaction is important for its apoptotic function.","method":"Colony formation assay, flow cytometry, immunofluorescence microscopy, deletion mutant analysis","journal":"Gastroenterology","confidence":"Medium","confidence_rationale":"Tier 2 — multiple orthogonal functional assays, single lab","pmids":["16012945"],"is_preprint":false},{"year":2007,"finding":"RASSF2 contains a functional bipartite nuclear localization signal (NLS); mutation of this NLS abolishes nuclear localization and diminishes its tumor suppressor/growth-suppressive activity, demonstrating that nuclear localization is required for function.","method":"NLS mutagenesis, immunofluorescence localization, in vitro and in vivo growth suppression assays","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 1-2 — mutagenesis combined with direct localization experiments and functional growth-suppression readouts","pmids":["17891178"],"is_preprint":false},{"year":2008,"finding":"A RASSF2 deletion mutant lacking the RA domain is unable to interact with Ras and exhibits less pro-apoptotic activity than full-length RASSF2, indicating the Ras-association domain is required for full pro-apoptotic activity in oral squamous cell carcinoma cells.","method":"Deletion mutant expression, apoptosis assays","journal":"Cancer science","confidence":"Medium","confidence_rationale":"Tier 2 — domain deletion mutagenesis with functional apoptosis readout, single lab","pmids":["18294275"],"is_preprint":false},{"year":2009,"finding":"RASSF2 associates with proapoptotic kinases MST1 and MST2 (confirmed at endogenous levels by co-immunoprecipitation), co-immunoprecipitates active MST1/2, is phosphorylated by co-immunoprecipitating MST1/2, and stabilizes MST2 by protecting it from proteolytic degradation. RASSF2 and MST2 colocalize; RASSF2 alone is nuclear but relocalizes to the cytoplasm in the presence of MST1 or MST2.","method":"Reciprocal co-immunoprecipitation at endogenous levels, stable and transient expression, siRNA knockdown, colocalization by immunofluorescence, kinase activity assay","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 2 — reciprocal endogenous Co-IP, multiple orthogonal methods including kinase assay and siRNA knockdown, replicated across cell lines","pmids":["19525978"],"is_preprint":false},{"year":2009,"finding":"MST1 kinase regulates RASSF2 protein stability (MST1 knockdown destabilizes RASSF2; Mst1-deficient mice show reduced Rassf2 protein). Conversely, RASSF2 activates MST1 kinase activity through formation of a RASSF2-MST1 complex, which inhibits the MST1-FOXO3 signaling pathway. RASSF2 also engages the JNK pathway to induce apoptosis in an MST1-independent manner.","method":"siRNA knockdown, complex formation assay, kinase activity assay, mouse knockout model, pathway analysis","journal":"Biochemical and biophysical research communications","confidence":"High","confidence_rationale":"Tier 2 — in vitro and in vivo (knockout mouse) evidence with multiple orthogonal methods and kinase activity assay","pmids":["19962960"],"is_preprint":false},{"year":2009,"finding":"RASSF2 contains a functional nuclear export signal (NES) in amino acids 240-260 (C-terminus); substitution of Ile254, Val257, Leu259 impairs nuclear export. Wild-type RASSF2 interacts with export receptor CRM-1 and is exported from the nucleus. ERK2 phosphorylates RASSF2, and MAPK pathway inhibition blocks RASSF2 phosphorylation and nuclear export. Nuclear import-defective RASSF2 fails to induce G1/S cell cycle arrest and apoptosis, demonstrating that nuclear localization is required for growth control.","method":"NES mutagenesis, leptomycin B treatment, CRM-1 co-immunoprecipitation, in vitro phosphorylation assay, MAPK inhibitors, cell cycle/apoptosis analysis","journal":"Experimental cell research","confidence":"High","confidence_rationale":"Tier 1-2 — mutagenesis, in vitro kinase assay, direct receptor interaction, functional readouts, multiple orthogonal methods","pmids":["19555684"],"is_preprint":false},{"year":2010,"finding":"RASSF2 forms a direct and endogenous complex with PAR-4 tumor suppressor; this interaction is regulated by K-Ras and is essential for the full apoptotic effects of PAR-4. RASSF2 modulates the nuclear translocation of PAR-4 from the cytoplasm to the nucleus in prostate tumor cells, providing a mechanism for its biological effects.","method":"Co-immunoprecipitation (endogenous), nuclear translocation assay, apoptosis assays with K-Ras regulation","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 2 — endogenous Co-IP, K-Ras regulation tested, functional nuclear translocation assay with apoptosis readout","pmids":["20368356"],"is_preprint":false},{"year":2010,"finding":"Deletion of the MST interaction domain of RASSF2 significantly reduces apoptosis induction in thyroid cancer cells, demonstrating that the MST1/2 interaction is required for RASSF2-mediated apoptosis.","method":"Domain deletion mutagenesis, apoptosis assay","journal":"Molecular cancer","confidence":"Medium","confidence_rationale":"Tier 2 — domain mutagenesis with quantified functional readout, single lab","pmids":["20920251"],"is_preprint":false},{"year":2012,"finding":"RASSF2 and K-Ras form an endogenous complex (validated by co-immunoprecipitation). Loss of RASSF2 expression in lung cancer cells with oncogenic K-Ras results in increased activated AKT levels, indicating that RASSF2 modulates Ras-AKT signaling. Loss of RASSF2 also confers resistance to taxol and cisplatin.","method":"Endogenous co-immunoprecipitation, RNAi knockdown, AKT phosphorylation assay, drug resistance assays","journal":"Molecular biology international","confidence":"Medium","confidence_rationale":"Tier 2 — endogenous Co-IP and RNAi with signaling readout, single lab","pmids":["22693671"],"is_preprint":false},{"year":2012,"finding":"Rassf2 knockout mice develop bone remodeling defects and hematopoietic anomalies. Rassf2 deficiency suppresses osteoblastogenesis but promotes osteoclastogenesis. RASSF2 associates with IKKα and IKKβ and suppresses IKK activity; Rassf2 deficiency results in NF-κB hyperactivation during osteoclast and osteoblast differentiation. Introduction of RASSF2 or dominant-negative IKK into Rassf2-/- precursors normalizes differentiation.","method":"Knockout mouse model, bone marrow transplantation, in vitro differentiation assays, co-immunoprecipitation, IKK activity assay, dominant-negative rescue","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 1-2 — in vivo knockout, in vitro reconstitution, IKK kinase assay, rescue experiments; multiple orthogonal methods","pmids":["22227519"],"is_preprint":false},{"year":2016,"finding":"Proteomics identified novel RASSF2 interaction partners including C1QBP, Vimentin, Protein phosphatase 1G, and Ribonuclease inhibitor. C1QBP interaction with RASSF2 is enhanced by K-Ras, whereas Vimentin interaction is reduced by K-Ras. RASSF2/K-Ras regulates the acetylation state of Vimentin.","method":"Proteomics (mass spectrometry), co-immunoprecipitation validation, acetylation analysis","journal":"Cancers","confidence":"Medium","confidence_rationale":"Tier 3 — MS-based interactome with Co-IP validation for two partners, single lab","pmids":["26999212"],"is_preprint":false},{"year":2017,"finding":"miR-7 targets RASSF2 in cancer-associated fibroblasts; overexpression of miR-7 leads to downregulation of RASSF2, which decreases PAR-4 secretion from fibroblasts and enhances cancer cell proliferation and migration in co-culture.","method":"miRNA overexpression/inhibition, co-culture assays, bioinformatics target validation, PAR-4 secretion measurement","journal":"Oncotarget","confidence":"Medium","confidence_rationale":"Tier 3 — functional co-culture assay with miRNA-RASSF2-PAR4 axis, single lab with multiple cell-based readouts","pmids":["27901488"],"is_preprint":false},{"year":2020,"finding":"RASSF2 re-expression in t(8;21) AML inhibits leukemia development in multiple models. RASSF2 functions depend on interaction with MST1 and MST2 (Hippo kinases) but are independent of canonical Hippo pathway signaling. Proximity-based biotin labeling (BioID) reveals RASSF2 associates with Rac GTPase-related proteins including the GEF DOCK2. RASSF2 knockdown impairs Rac GTPase activation.","method":"Re-expression in AML models, BioID proximity labeling, co-immunoprecipitation, Rac GTPase activation assay, MST1/2 interaction studies","journal":"Blood cancer journal","confidence":"High","confidence_rationale":"Tier 1-2 — proximity labeling proteomics, direct GTPase activity assay, functional in vivo and in vitro models, multiple orthogonal methods","pmids":["32029705"],"is_preprint":false},{"year":2026,"finding":"Cell-surface nucleolin (NCL) interacts with RASSF2 via its RNA-binding domain and facilitates nuclear transport of RASSF2, thereby promoting endothelial cell pyroptosis and inflammatory responses in atherosclerosis; NCL suppression decreases nuclear RASSF2 expression.","method":"Immunoprecipitation-mass spectrometry, siRNA knockdown, nuclear fractionation, pyroptosis assays in ApoE-/- mouse model","journal":"Atherosclerosis","confidence":"Medium","confidence_rationale":"Tier 2 — IP-MS with functional validation in mouse model, single lab","pmids":["41895182"],"is_preprint":false}],"current_model":"RASSF2 is a K-Ras-specific effector and tumor suppressor that binds K-Ras in a GTP-dependent manner; it shuttles between nucleus and cytoplasm via functional NLS and NES sequences (the latter regulated by ERK2 phosphorylation), and nuclear localization is required for its pro-apoptotic and cell cycle arrest functions; in the nucleus it forms endogenous complexes with MST1/MST2 (stabilizing these kinases and activating their activity) and with PAR-4 (facilitating PAR-4 nuclear translocation to promote apoptosis); it suppresses NF-κB signaling by binding and inhibiting IKKα/β to regulate osteoblast and osteoclast differentiation; and it associates with Rac GTPase pathway components (including DOCK2) to support Rac activation, with additional partners including C1QBP and Vimentin whose interactions are modulated by K-Ras."},"narrative":{"teleology":[{"year":2003,"claim":"The identity of RASSF2 as a Ras effector was unknown; GTP-dependent binding assays established it as a K-Ras-specific interactor that induces apoptosis and cell cycle arrest, framing it as a tumor suppressor downstream of Ras.","evidence":"Direct binding assay demonstrating GTP-dependent K-Ras interaction with selectivity over H-Ras, plus overexpression-based apoptosis/cell cycle readouts","pmids":["12732644"],"confidence":"High","gaps":["No structural basis for K-Ras selectivity","Endogenous complex not yet validated","Downstream signaling pathway not defined"]},{"year":2005,"claim":"Whether the RA domain was functionally required for tumor suppression was untested; deletion mutant analysis in colorectal and oral cancer cells showed that the RA domain is necessary for full pro-apoptotic activity, linking Ras binding to the apoptotic mechanism.","evidence":"Colony formation, flow cytometry, and deletion mutant analysis in colorectal and oral squamous cell carcinoma lines","pmids":["16012945","18294275"],"confidence":"Medium","gaps":["Mechanism connecting Ras binding to apoptosis execution not identified","RA-domain deletion may affect protein folding rather than interaction specifically"]},{"year":2007,"claim":"The subcellular site of RASSF2 action was unclear; identification of a functional bipartite NLS and demonstration that its mutation abolishes growth-suppressive activity established nuclear localization as essential for tumor suppressor function.","evidence":"NLS mutagenesis with immunofluorescence and in vitro/in vivo growth suppression assays","pmids":["17891178"],"confidence":"High","gaps":["Nuclear targets mediating growth suppression not identified","Regulation of nuclear import not addressed"]},{"year":2009,"claim":"How RASSF2 nucleocytoplasmic shuttling is regulated was unknown; identification of a functional NES (aa 240–260) and demonstration that ERK2 phosphorylation drives CRM-1-dependent nuclear export revealed that the MAPK pathway antagonizes RASSF2 nuclear function.","evidence":"NES mutagenesis, leptomycin B treatment, CRM-1 Co-IP, in vitro ERK2 kinase assay, MAPK inhibitor experiments","pmids":["19555684"],"confidence":"High","gaps":["Specific ERK2 phosphorylation sites not fully mapped","Physiological contexts triggering RASSF2 nuclear export not defined"]},{"year":2009,"claim":"The effector pathway linking RASSF2 to apoptosis execution was undefined; reciprocal endogenous co-immunoprecipitation and kinase assays showed that RASSF2 forms complexes with MST1/MST2, stabilizes MST2 against degradation, and activates MST1 kinase activity, establishing the Hippo kinase axis as a core RASSF2 effector mechanism.","evidence":"Endogenous reciprocal Co-IP, siRNA knockdown, Mst1-knockout mouse, in vitro kinase activity assay, colocalization microscopy","pmids":["19525978","19962960"],"confidence":"High","gaps":["Whether RASSF2 activates canonical Hippo (YAP/TAZ) signaling downstream of MST not resolved","Structural basis of RASSF2-MST interaction not determined"]},{"year":2010,"claim":"Whether RASSF2 cooperates with other tumor suppressors was open; endogenous complex formation with PAR-4 and demonstration that RASSF2 facilitates K-Ras-dependent PAR-4 nuclear translocation identified a second apoptotic effector arm distinct from MST.","evidence":"Endogenous Co-IP, nuclear translocation assay, apoptosis assays with K-Ras modulation in prostate cancer cells","pmids":["20368356"],"confidence":"High","gaps":["Whether MST and PAR-4 arms are redundant or synergistic not tested","Direct versus indirect nature of K-Ras regulation of RASSF2-PAR-4 not distinguished"]},{"year":2010,"claim":"Whether the MST interaction domain is functionally required was not directly tested by domain deletion in the apoptosis context; deletion of the MST-binding region significantly reduced apoptosis induction in thyroid cancer cells, confirming functional dependence.","evidence":"Domain deletion mutagenesis with apoptosis quantification in thyroid cancer cells","pmids":["20920251"],"confidence":"Medium","gaps":["Domain deletion may disrupt other interactions besides MST","Downstream MST substrates mediating the apoptotic signal not identified"]},{"year":2012,"claim":"The physiological role of RASSF2 beyond cancer was unknown; Rassf2-knockout mice revealed bone remodeling defects due to NF-κB hyperactivation, and biochemical studies showed RASSF2 directly binds and inhibits IKKα/IKKβ, establishing a non-Hippo effector pathway controlling osteoblast/osteoclast differentiation.","evidence":"Knockout mouse, bone marrow transplantation, in vitro differentiation, IKK activity assay, Co-IP, dominant-negative IKK rescue","pmids":["22227519"],"confidence":"High","gaps":["Whether IKK inhibition contributes to RASSF2 tumor suppressor activity not tested","Structural basis of IKK inhibition unknown"]},{"year":2012,"claim":"Whether RASSF2 modulates canonical Ras effector pathways was unclear; endogenous RASSF2-K-Ras co-immunoprecipitation and knockdown experiments demonstrated that RASSF2 loss increases activated AKT levels, linking RASSF2 to restraint of PI3K-AKT signaling.","evidence":"Endogenous Co-IP, RNAi knockdown with AKT phosphorylation readout in K-Ras-mutant lung cancer cells","pmids":["22693671"],"confidence":"Medium","gaps":["Mechanism by which RASSF2 suppresses AKT activation not defined","Direct versus indirect effect on PI3K pathway not resolved"]},{"year":2020,"claim":"Whether RASSF2 engages pathways beyond Hippo and NF-κB was unresolved; BioID proximity labeling in AML cells revealed association with Rac GTPase pathway components including the GEF DOCK2, and RASSF2 knockdown impaired Rac activation, identifying a new signaling axis through which RASSF2 suppresses leukemia.","evidence":"BioID proximity labeling, Co-IP, Rac GTPase activation assay, re-expression in multiple AML models","pmids":["32029705"],"confidence":"High","gaps":["Direct versus adaptor role for RASSF2 in Rac activation not distinguished","Whether DOCK2-Rac axis operates in solid tumors not tested"]},{"year":2026,"claim":"How RASSF2 nuclear import is regulated by extracellular cues was unknown; identification of cell-surface nucleolin as a binding partner that facilitates RASSF2 nuclear transport expanded the import mechanism and linked RASSF2 nuclear accumulation to endothelial pyroptosis in atherosclerosis.","evidence":"IP-mass spectrometry, siRNA knockdown of nucleolin, nuclear fractionation, pyroptosis assays in ApoE-/- mouse model","pmids":["41895182"],"confidence":"Medium","gaps":["Mechanism by which surface nucleolin accesses cytoplasmic RASSF2 not explained","Pyroptosis pathway downstream of nuclear RASSF2 not defined","Single lab; independent replication needed"]},{"year":null,"claim":"The structural basis for K-Ras selectivity, the precise mechanism by which RASSF2 inhibits IKK, and whether the MST, PAR-4, Rac/DOCK2, and NF-κB effector arms are coordinated or context-specific remain unresolved.","evidence":"","pmids":[],"confidence":"Low","gaps":["No crystal or cryo-EM structure of RASSF2 or any of its complexes","Integration of MST, PAR-4, IKK, and Rac pathways not tested in a single system","In vivo tumor suppressor validation limited to Rassf2-knockout bone phenotype; no tumor model published"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[4,5,10]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,7,13]}],"localization":[{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[2,6,7,14]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[4,6]}],"pathway":[{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[0,1,3,4,5,7,8]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,9,10,13]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[10]}],"complexes":[],"partners":["KRAS","MST1","MST2","PAWR","IKBKA","IKBKB","DOCK2","NCL"],"other_free_text":[]},"mechanistic_narrative":"RASSF2 is a K-Ras-specific effector and tumor suppressor that integrates Ras signaling with apoptosis, cell cycle control, and NF-κB-dependent differentiation programs. It binds K-Ras in a GTP-dependent manner through its Ras-association (RA) domain and shuttles between cytoplasm and nucleus via a bipartite NLS and an ERK2-phosphorylation-regulated NES, with nuclear localization required for its growth-suppressive and pro-apoptotic functions [PMID:12732644, PMID:17891178, PMID:19555684]. In the nucleus, RASSF2 forms endogenous complexes with the Hippo pathway kinases MST1/MST2—stabilizing MST2, activating MST1 kinase activity, and requiring this interaction for full apoptosis induction—and with the tumor suppressor PAR-4, facilitating PAR-4 nuclear translocation to promote apoptosis [PMID:19525978, PMID:19962960, PMID:20368356]. RASSF2 also suppresses NF-κB signaling by directly binding and inhibiting IKKα/IKKβ, thereby regulating osteoblast and osteoclast differentiation in vivo, and associates with the Rac GEF DOCK2 to support Rac GTPase activation in hematopoietic cells [PMID:22227519, PMID:32029705]."},"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":150,"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":113,"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":69,"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":61,"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":"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":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":46,"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":"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":42,"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 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Partners.","date":"2016","source":"Cancers","url":"https://pubmed.ncbi.nlm.nih.gov/26999212","citation_count":9,"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":1,"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":1,"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}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":14318,"output_tokens":3700,"usd":0.049227},"stage2":{"model":"claude-opus-4-6","input_tokens":7122,"output_tokens":3052,"usd":0.167865},"total_usd":0.217092,"stage1_batch_id":"msgbatch_01SUR8NDzVrNmyszWyJeCct7","stage2_batch_id":"msgbatch_01G7Yfutko3cDWzejrwH5AMx","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, identifying it as a K-Ras-specific effector that promotes apoptosis and cell cycle arrest.\",\n      \"method\": \"Direct binding assay (GTP-dependent interaction), overexpression with apoptosis/cell cycle readouts\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — direct binding assay with GTP-dependence specificity, replicated by multiple subsequent studies\",\n      \"pmids\": [\"12732644\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"RASSF2 expression induces morphological changes and apoptosis in colorectal cancer cells; a deletion mutant lacking the RA domain shows reduced pro-apoptotic activity, indicating the Ras interaction is important for its apoptotic function.\",\n      \"method\": \"Colony formation assay, flow cytometry, immunofluorescence microscopy, deletion mutant analysis\",\n      \"journal\": \"Gastroenterology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal functional assays, 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 this NLS abolishes nuclear localization and diminishes its tumor suppressor/growth-suppressive activity, demonstrating that nuclear localization is required for function.\",\n      \"method\": \"NLS mutagenesis, immunofluorescence localization, in vitro and in vivo growth suppression assays\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — mutagenesis combined with direct localization experiments and functional growth-suppression readouts\",\n      \"pmids\": [\"17891178\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"A RASSF2 deletion mutant lacking the RA domain is unable to interact with Ras and exhibits less pro-apoptotic activity than full-length RASSF2, indicating the Ras-association domain is required for full pro-apoptotic activity in oral squamous cell carcinoma cells.\",\n      \"method\": \"Deletion mutant expression, apoptosis assays\",\n      \"journal\": \"Cancer science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — domain deletion mutagenesis with functional apoptosis readout, single lab\",\n      \"pmids\": [\"18294275\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"RASSF2 associates with proapoptotic kinases MST1 and MST2 (confirmed at endogenous levels by co-immunoprecipitation), co-immunoprecipitates active MST1/2, is phosphorylated by co-immunoprecipitating MST1/2, and stabilizes MST2 by protecting it from proteolytic degradation. RASSF2 and MST2 colocalize; RASSF2 alone is nuclear but relocalizes to the cytoplasm in the presence of MST1 or MST2.\",\n      \"method\": \"Reciprocal co-immunoprecipitation at endogenous levels, stable and transient expression, siRNA knockdown, colocalization by immunofluorescence, kinase activity assay\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal endogenous Co-IP, multiple orthogonal methods including kinase assay and siRNA knockdown, replicated across cell lines\",\n      \"pmids\": [\"19525978\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"MST1 kinase regulates RASSF2 protein stability (MST1 knockdown destabilizes RASSF2; Mst1-deficient mice show reduced Rassf2 protein). Conversely, RASSF2 activates MST1 kinase activity through formation of a RASSF2-MST1 complex, which inhibits the MST1-FOXO3 signaling pathway. RASSF2 also engages the JNK pathway to induce apoptosis in an MST1-independent manner.\",\n      \"method\": \"siRNA knockdown, complex formation assay, kinase activity assay, mouse knockout model, pathway analysis\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — in vitro and in vivo (knockout mouse) evidence with multiple orthogonal methods and kinase activity assay\",\n      \"pmids\": [\"19962960\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"RASSF2 contains a functional nuclear export signal (NES) in amino acids 240-260 (C-terminus); substitution of Ile254, Val257, Leu259 impairs nuclear export. Wild-type RASSF2 interacts with export receptor CRM-1 and is exported from the nucleus. ERK2 phosphorylates RASSF2, and MAPK pathway inhibition blocks RASSF2 phosphorylation and nuclear export. Nuclear import-defective RASSF2 fails to induce G1/S cell cycle arrest and apoptosis, demonstrating that nuclear localization is required for growth control.\",\n      \"method\": \"NES mutagenesis, leptomycin B treatment, CRM-1 co-immunoprecipitation, in vitro phosphorylation assay, MAPK inhibitors, cell cycle/apoptosis analysis\",\n      \"journal\": \"Experimental cell research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — mutagenesis, in vitro kinase assay, direct receptor interaction, functional readouts, multiple orthogonal methods\",\n      \"pmids\": [\"19555684\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"RASSF2 forms a direct and endogenous complex with PAR-4 tumor suppressor; this interaction is regulated by K-Ras and is essential for the full apoptotic effects of PAR-4. RASSF2 modulates the nuclear translocation of PAR-4 from the cytoplasm to the nucleus in prostate tumor cells, providing a mechanism for its biological effects.\",\n      \"method\": \"Co-immunoprecipitation (endogenous), nuclear translocation assay, apoptosis assays with K-Ras regulation\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — endogenous Co-IP, K-Ras regulation tested, functional nuclear translocation assay with apoptosis readout\",\n      \"pmids\": [\"20368356\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Deletion of the MST interaction domain of RASSF2 significantly reduces apoptosis induction in thyroid cancer cells, demonstrating that the MST1/2 interaction is required for RASSF2-mediated apoptosis.\",\n      \"method\": \"Domain deletion mutagenesis, apoptosis assay\",\n      \"journal\": \"Molecular cancer\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — domain mutagenesis with quantified functional readout, single lab\",\n      \"pmids\": [\"20920251\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"RASSF2 and K-Ras form an endogenous complex (validated by co-immunoprecipitation). Loss of RASSF2 expression in lung cancer cells with oncogenic K-Ras results in increased activated AKT levels, indicating that RASSF2 modulates Ras-AKT signaling. Loss of RASSF2 also confers resistance to taxol and cisplatin.\",\n      \"method\": \"Endogenous co-immunoprecipitation, RNAi knockdown, AKT phosphorylation assay, drug resistance assays\",\n      \"journal\": \"Molecular biology international\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — endogenous Co-IP and RNAi with signaling readout, single lab\",\n      \"pmids\": [\"22693671\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Rassf2 knockout mice develop bone remodeling defects and hematopoietic anomalies. Rassf2 deficiency suppresses osteoblastogenesis but promotes osteoclastogenesis. RASSF2 associates with IKKα and IKKβ and suppresses IKK activity; Rassf2 deficiency results in NF-κB hyperactivation during osteoclast and osteoblast differentiation. Introduction of RASSF2 or dominant-negative IKK into Rassf2-/- precursors normalizes differentiation.\",\n      \"method\": \"Knockout mouse model, bone marrow transplantation, in vitro differentiation assays, co-immunoprecipitation, IKK activity assay, dominant-negative rescue\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — in vivo knockout, in vitro reconstitution, IKK kinase assay, rescue experiments; multiple orthogonal methods\",\n      \"pmids\": [\"22227519\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Proteomics identified novel RASSF2 interaction partners including C1QBP, Vimentin, Protein phosphatase 1G, and Ribonuclease inhibitor. C1QBP interaction with RASSF2 is enhanced by K-Ras, whereas Vimentin interaction is reduced by K-Ras. RASSF2/K-Ras regulates the acetylation state of Vimentin.\",\n      \"method\": \"Proteomics (mass spectrometry), co-immunoprecipitation validation, acetylation analysis\",\n      \"journal\": \"Cancers\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — MS-based interactome with Co-IP validation for two partners, single lab\",\n      \"pmids\": [\"26999212\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"miR-7 targets RASSF2 in cancer-associated fibroblasts; overexpression of miR-7 leads to downregulation of RASSF2, which decreases PAR-4 secretion from fibroblasts and enhances cancer cell proliferation and migration in co-culture.\",\n      \"method\": \"miRNA overexpression/inhibition, co-culture assays, bioinformatics target validation, PAR-4 secretion measurement\",\n      \"journal\": \"Oncotarget\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — functional co-culture assay with miRNA-RASSF2-PAR4 axis, single lab with multiple cell-based readouts\",\n      \"pmids\": [\"27901488\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"RASSF2 re-expression in t(8;21) AML inhibits leukemia development in multiple models. RASSF2 functions depend on interaction with MST1 and MST2 (Hippo kinases) but are independent of canonical Hippo pathway signaling. Proximity-based biotin labeling (BioID) reveals RASSF2 associates with Rac GTPase-related proteins including the GEF DOCK2. RASSF2 knockdown impairs Rac GTPase activation.\",\n      \"method\": \"Re-expression in AML models, BioID proximity labeling, co-immunoprecipitation, Rac GTPase activation assay, MST1/2 interaction studies\",\n      \"journal\": \"Blood cancer journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — proximity labeling proteomics, direct GTPase activity assay, functional in vivo and in vitro models, multiple orthogonal methods\",\n      \"pmids\": [\"32029705\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"Cell-surface nucleolin (NCL) interacts with RASSF2 via its RNA-binding domain and facilitates nuclear transport of RASSF2, thereby promoting endothelial cell pyroptosis and inflammatory responses in atherosclerosis; NCL suppression decreases nuclear RASSF2 expression.\",\n      \"method\": \"Immunoprecipitation-mass spectrometry, siRNA knockdown, nuclear fractionation, pyroptosis assays in ApoE-/- mouse model\",\n      \"journal\": \"Atherosclerosis\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — IP-MS with functional validation in mouse model, single lab\",\n      \"pmids\": [\"41895182\"],\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; it shuttles between nucleus and cytoplasm via functional NLS and NES sequences (the latter regulated by ERK2 phosphorylation), and nuclear localization is required for its pro-apoptotic and cell cycle arrest functions; in the nucleus it forms endogenous complexes with MST1/MST2 (stabilizing these kinases and activating their activity) and with PAR-4 (facilitating PAR-4 nuclear translocation to promote apoptosis); it suppresses NF-κB signaling by binding and inhibiting IKKα/β to regulate osteoblast and osteoclast differentiation; and it associates with Rac GTPase pathway components (including DOCK2) to support Rac activation, with additional partners including C1QBP and Vimentin whose interactions are modulated by K-Ras.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"RASSF2 is a K-Ras-specific effector and tumor suppressor that integrates Ras signaling with apoptosis, cell cycle control, and NF-κB-dependent differentiation programs. It binds K-Ras in a GTP-dependent manner through its Ras-association (RA) domain and shuttles between cytoplasm and nucleus via a bipartite NLS and an ERK2-phosphorylation-regulated NES, with nuclear localization required for its growth-suppressive and pro-apoptotic functions [PMID:12732644, PMID:17891178, PMID:19555684]. In the nucleus, RASSF2 forms endogenous complexes with the Hippo pathway kinases MST1/MST2—stabilizing MST2, activating MST1 kinase activity, and requiring this interaction for full apoptosis induction—and with the tumor suppressor PAR-4, facilitating PAR-4 nuclear translocation to promote apoptosis [PMID:19525978, PMID:19962960, PMID:20368356]. RASSF2 also suppresses NF-κB signaling by directly binding and inhibiting IKKα/IKKβ, thereby regulating osteoblast and osteoclast differentiation in vivo, and associates with the Rac GEF DOCK2 to support Rac GTPase activation in hematopoietic cells [PMID:22227519, PMID:32029705].\",\n  \"teleology\": [\n    {\n      \"year\": 2003,\n      \"claim\": \"The identity of RASSF2 as a Ras effector was unknown; GTP-dependent binding assays established it as a K-Ras-specific interactor that induces apoptosis and cell cycle arrest, framing it as a tumor suppressor downstream of Ras.\",\n      \"evidence\": \"Direct binding assay demonstrating GTP-dependent K-Ras interaction with selectivity over H-Ras, plus overexpression-based apoptosis/cell cycle readouts\",\n      \"pmids\": [\"12732644\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No structural basis for K-Ras selectivity\", \"Endogenous complex not yet validated\", \"Downstream signaling pathway not defined\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Whether the RA domain was functionally required for tumor suppression was untested; deletion mutant analysis in colorectal and oral cancer cells showed that the RA domain is necessary for full pro-apoptotic activity, linking Ras binding to the apoptotic mechanism.\",\n      \"evidence\": \"Colony formation, flow cytometry, and deletion mutant analysis in colorectal and oral squamous cell carcinoma lines\",\n      \"pmids\": [\"16012945\", \"18294275\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism connecting Ras binding to apoptosis execution not identified\", \"RA-domain deletion may affect protein folding rather than interaction specifically\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"The subcellular site of RASSF2 action was unclear; identification of a functional bipartite NLS and demonstration that its mutation abolishes growth-suppressive activity established nuclear localization as essential for tumor suppressor function.\",\n      \"evidence\": \"NLS mutagenesis with immunofluorescence and in vitro/in vivo growth suppression assays\",\n      \"pmids\": [\"17891178\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Nuclear targets mediating growth suppression not identified\", \"Regulation of nuclear import not addressed\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"How RASSF2 nucleocytoplasmic shuttling is regulated was unknown; identification of a functional NES (aa 240–260) and demonstration that ERK2 phosphorylation drives CRM-1-dependent nuclear export revealed that the MAPK pathway antagonizes RASSF2 nuclear function.\",\n      \"evidence\": \"NES mutagenesis, leptomycin B treatment, CRM-1 Co-IP, in vitro ERK2 kinase assay, MAPK inhibitor experiments\",\n      \"pmids\": [\"19555684\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Specific ERK2 phosphorylation sites not fully mapped\", \"Physiological contexts triggering RASSF2 nuclear export not defined\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"The effector pathway linking RASSF2 to apoptosis execution was undefined; reciprocal endogenous co-immunoprecipitation and kinase assays showed that RASSF2 forms complexes with MST1/MST2, stabilizes MST2 against degradation, and activates MST1 kinase activity, establishing the Hippo kinase axis as a core RASSF2 effector mechanism.\",\n      \"evidence\": \"Endogenous reciprocal Co-IP, siRNA knockdown, Mst1-knockout mouse, in vitro kinase activity assay, colocalization microscopy\",\n      \"pmids\": [\"19525978\", \"19962960\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether RASSF2 activates canonical Hippo (YAP/TAZ) signaling downstream of MST not resolved\", \"Structural basis of RASSF2-MST interaction not determined\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Whether RASSF2 cooperates with other tumor suppressors was open; endogenous complex formation with PAR-4 and demonstration that RASSF2 facilitates K-Ras-dependent PAR-4 nuclear translocation identified a second apoptotic effector arm distinct from MST.\",\n      \"evidence\": \"Endogenous Co-IP, nuclear translocation assay, apoptosis assays with K-Ras modulation in prostate cancer cells\",\n      \"pmids\": [\"20368356\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether MST and PAR-4 arms are redundant or synergistic not tested\", \"Direct versus indirect nature of K-Ras regulation of RASSF2-PAR-4 not distinguished\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Whether the MST interaction domain is functionally required was not directly tested by domain deletion in the apoptosis context; deletion of the MST-binding region significantly reduced apoptosis induction in thyroid cancer cells, confirming functional dependence.\",\n      \"evidence\": \"Domain deletion mutagenesis with apoptosis quantification in thyroid cancer cells\",\n      \"pmids\": [\"20920251\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Domain deletion may disrupt other interactions besides MST\", \"Downstream MST substrates mediating the apoptotic signal not identified\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"The physiological role of RASSF2 beyond cancer was unknown; Rassf2-knockout mice revealed bone remodeling defects due to NF-κB hyperactivation, and biochemical studies showed RASSF2 directly binds and inhibits IKKα/IKKβ, establishing a non-Hippo effector pathway controlling osteoblast/osteoclast differentiation.\",\n      \"evidence\": \"Knockout mouse, bone marrow transplantation, in vitro differentiation, IKK activity assay, Co-IP, dominant-negative IKK rescue\",\n      \"pmids\": [\"22227519\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether IKK inhibition contributes to RASSF2 tumor suppressor activity not tested\", \"Structural basis of IKK inhibition unknown\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Whether RASSF2 modulates canonical Ras effector pathways was unclear; endogenous RASSF2-K-Ras co-immunoprecipitation and knockdown experiments demonstrated that RASSF2 loss increases activated AKT levels, linking RASSF2 to restraint of PI3K-AKT signaling.\",\n      \"evidence\": \"Endogenous Co-IP, RNAi knockdown with AKT phosphorylation readout in K-Ras-mutant lung cancer cells\",\n      \"pmids\": [\"22693671\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism by which RASSF2 suppresses AKT activation not defined\", \"Direct versus indirect effect on PI3K pathway not resolved\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Whether RASSF2 engages pathways beyond Hippo and NF-κB was unresolved; BioID proximity labeling in AML cells revealed association with Rac GTPase pathway components including the GEF DOCK2, and RASSF2 knockdown impaired Rac activation, identifying a new signaling axis through which RASSF2 suppresses leukemia.\",\n      \"evidence\": \"BioID proximity labeling, Co-IP, Rac GTPase activation assay, re-expression in multiple AML models\",\n      \"pmids\": [\"32029705\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct versus adaptor role for RASSF2 in Rac activation not distinguished\", \"Whether DOCK2-Rac axis operates in solid tumors not tested\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"How RASSF2 nuclear import is regulated by extracellular cues was unknown; identification of cell-surface nucleolin as a binding partner that facilitates RASSF2 nuclear transport expanded the import mechanism and linked RASSF2 nuclear accumulation to endothelial pyroptosis in atherosclerosis.\",\n      \"evidence\": \"IP-mass spectrometry, siRNA knockdown of nucleolin, nuclear fractionation, pyroptosis assays in ApoE-/- mouse model\",\n      \"pmids\": [\"41895182\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism by which surface nucleolin accesses cytoplasmic RASSF2 not explained\", \"Pyroptosis pathway downstream of nuclear RASSF2 not defined\", \"Single lab; independent replication needed\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The structural basis for K-Ras selectivity, the precise mechanism by which RASSF2 inhibits IKK, and whether the MST, PAR-4, Rac/DOCK2, and NF-κB effector arms are coordinated or context-specific remain unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No crystal or cryo-EM structure of RASSF2 or any of its complexes\", \"Integration of MST, PAR-4, IKK, and Rac pathways not tested in a single system\", \"In vivo tumor suppressor validation limited to Rassf2-knockout bone phenotype; no tumor model published\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [4, 5, 10]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 7, 13]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [2, 6, 7, 14]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [4, 6]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [0, 1, 3, 4, 5, 7, 8]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 9, 10, 13]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [10]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"KRAS\", \"MST1\", \"MST2\", \"PAWR\", \"IKBKA\", \"IKBKB\", \"DOCK2\", \"NCL\"],\n    \"other_free_text\": []\n  }\n}\n```"}