{"gene":"RASSF6","run_date":"2026-06-10T06:43:36","timeline":{"discoveries":[{"year":2007,"finding":"RASSF6 possesses a Ras Association domain and binds activated Ras, promoting apoptosis and synergizing with activated K-Ras to induce cell death; suppression of RASSF6 enhanced the tumorigenic phenotype of a human lung tumor cell line.","method":"Exogenous expression, suppression experiments in tumor cell lines, cell death assays","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — functional KD/OE with defined phenotype and Ras-binding characterization in single lab","pmids":["17404571"],"is_preprint":false},{"year":2009,"finding":"RASSF6 binds MST2 (mammalian Hippo kinase homolog) and inhibits MST2 kinase activity, antagonizing Hippo signaling; upon MST2 activation, RASSF6 is released and causes apoptosis in a WW45 (mammalian Salvador homolog)-dependent manner, defining a RASSF6-mediated apoptotic pathway parallel to the canonical Hippo pathway.","method":"Co-immunoprecipitation, kinase activity assays, epistasis analysis with WW45","journal":"Science signaling","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal Co-IP, kinase assays, and genetic epistasis in single lab with multiple orthogonal methods","pmids":["19797269"],"is_preprint":false},{"year":2012,"finding":"RASSF6 binds PDZ domains of MAGI-1 through its C-terminal PDZ-binding motif and co-immunoprecipitates with MAGI-1 from rat liver; RASSF6 is localized in normal kidney glomerulus and on apical membranes of renal proximal tubular epithelial cells, and is involved in sorbitol-induced apoptosis together with the Hippo pathway in HK-2 cells.","method":"Yeast two-hybrid screening (original identification), co-immunoprecipitation from rat liver, immunolocalization in kidney, functional apoptosis assays in HK-2 cells","journal":"Journal of biochemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP from tissue, localization, and functional assay in single lab with multiple orthogonal methods","pmids":["22577168"],"is_preprint":false},{"year":2013,"finding":"RASSF6 binds MDM2 and facilitates its ubiquitination, thereby blocking MDM2-mediated p53 degradation and stabilizing p53; RASSF6 overexpression induces G1/S arrest in p53-positive cells, and its depletion prevents UV- and VP-16-induced apoptosis and G1/S arrest, delays DNA repair, and increases polyploid cells.","method":"Co-immunoprecipitation, ubiquitination assays, siRNA knockdown, cell cycle analysis, p53 target gene expression","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Moderate — Co-IP, ubiquitination assay, KD with multiple defined phenotypic readouts in single lab","pmids":["24003224"],"is_preprint":false},{"year":2013,"finding":"RASSF6 mRNA in adipocytes is downregulated by conditioned medium from activated macrophages, and decreased RASSF6 expression upregulates CD44 and HMGA2 mRNA expression in adipocytes.","method":"Gene expression profiling in 3T3-L1 adipocytes co-cultured with RAW264.7 macrophages, in vivo correlation with macrophage number","journal":"PloS one","confidence":"Low","confidence_rationale":"Tier 3 / Weak — expression-level findings with downstream gene targets noted, single lab, no direct mechanistic pathway dissection","pmids":["23626755"],"is_preprint":false},{"year":2014,"finding":"RASSF6 activates JNK signaling to promote apoptosis and p21(Cip1/Waf1)-dependent G1 cell cycle arrest in clear cell renal cell carcinoma cells; JNK inhibitor SP600125 blocked RASSF6-mediated apoptosis sensitization.","method":"Ectopic re-expression and siRNA knockdown, JNK inhibitor treatment, cell cycle analysis, tumor xenograft models","journal":"Cell cycle (Georgetown, Tex.)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — KD/OE with pharmacological inhibition of JNK and multiple phenotypic readouts, single lab","pmids":["24626183"],"is_preprint":false},{"year":2014,"finding":"RASSF6 re-expression in metastatic melanoma cells with B-Raf(V600E) inhibited invasion through collagen, suppressed MAPK pathway activation and AKT, and increased association of mutant B-Raf with MST1.","method":"Invasion assay through collagen, western blot for MAPK/AKT, co-immunoprecipitation of B-Raf and MST1","journal":"Epigenetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, functional invasion assay, and signaling readouts in single lab","pmids":["25482183"],"is_preprint":false},{"year":2016,"finding":"miR-181a-5p directly represses RASSF6 expression and thereby activates the MAPK signaling pathway, promoting gastric cancer cell proliferation, invasion, and EMT; overexpression or silencing of RASSF6 partially reverses the oncogenic effects of miR-181a-5p.","method":"Luciferase reporter assay for direct miR-181a-5p binding to RASSF6 3'UTR, miRNA mimic/inhibitor with rescue by RASSF6 OE/KD, in vitro and in vivo assays","journal":"Cancer letters","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — luciferase reporter (direct binding validation), rescue experiments, in vivo confirmation, single lab","pmids":["28043911"],"is_preprint":false},{"year":2016,"finding":"RASSF6 overexpression in HCC cells suppresses FAK phosphorylation and decreases MMP-2 and MMP-9 expression, inhibiting invasion and migration.","method":"Overexpression in HepG2 cells, western blot for p-FAK, MMP-2, MMP-9, invasion assays, xenograft model","journal":"Oncology research","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, OE only with western blot readouts, no direct mechanistic dissection of FAK regulation","pmids":["27983932"],"is_preprint":false},{"year":2017,"finding":"BCL-XL directly interacts with RASSF6, inhibits the interaction between RASSF6 and MDM2, and suppresses p53 expression, thereby antagonizing RASSF6-mediated apoptosis.","method":"Co-immunoprecipitation, p53 expression assays, apoptosis assays with BCL-XL manipulation","journal":"Genes to cells : devoted to molecular & cellular mechanisms","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP showing direct interaction and functional consequence on p53/apoptosis, single lab","pmids":["29193479"],"is_preprint":false},{"year":2018,"finding":"RASSF6 mediates cell cycle arrest and apoptosis via pRb independent of p53: RASSF6 enhances the interaction between pRb and protein phosphatase, suppresses BMI1 to increase P16INK4A and P14ARF expression, increases unphosphorylated pRb, augments pRb-E2F1 interaction, and induces TP73 target genes in a p53-negative background.","method":"Co-immunoprecipitation, western blot, siRNA knockdown, flow cytometry cell cycle analysis in p53-negative cells","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (Co-IP, gene expression, cell cycle), p53-negative background controls, multiple pathway nodes tested in single rigorous study","pmids":["29891515"],"is_preprint":false},{"year":2018,"finding":"RASSF6 activates the JNK signaling pathway, which contributes to Mcl-1 inhibition; pharmacological JNK pathway suppression partially restores Mcl-1 expression and sorafenib resistance, establishing a RASSF6-JNK-Mcl-1 axis.","method":"Stable RASSF6 overexpression in RCC cell lines, JNK inhibitor treatment, Mcl-1 expression assays, in vitro and in vivo sorafenib resistance assays","journal":"Cancer letters","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — stable OE, pharmacological rescue, in vitro and in vivo validation, single lab","pmids":["29864454"],"is_preprint":false},{"year":2018,"finding":"RASSF6 activates Hippo signaling by upregulating MST1/2 and LATS1 phosphorylation, represses YAP, and YAP repression mediates RASSF6's effects on Bcl-xL, apoptosis (cleaved caspase-3, cytochrome c), and mitochondrial membrane potential in breast cancer cells.","method":"Overexpression/depletion of RASSF6, YAP rescue experiments, western blot, JC-1 staining, flow cytometry","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — YAP rescue epistasis, multiple pathway markers, single lab","pmids":["29964010"],"is_preprint":false},{"year":2019,"finding":"miR-496 directly binds RASSF6 and represses its expression, activating Wnt signaling to promote CRC cell migration and EMT; this effect was validated by luciferase reporter assay and RASSF6 siRNA rescue experiments.","method":"Luciferase reporter assay, miR-496 mimic/inhibitor, RASSF6 siRNA rescue, migration and EMT assays","journal":"Journal of cellular physiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct binding validated by luciferase reporter, functional rescue, single lab","pmids":["31273789"],"is_preprint":false},{"year":2019,"finding":"RASSF6 overexpression in bladder cancer cells downregulates YAP (Hippo pathway effector) and Bcl-xL; YAP silencing abolishes the effects of RASSF6 on Bcl-xL expression, doxorubicin-induced apoptosis, and mitochondrial membrane potential, placing YAP downstream of RASSF6.","method":"Overexpression/siRNA knockdown, YAP rescue/epistasis, JC-1 staining, western blot, AnnexinV/PI apoptosis assay","journal":"OncoTargets and therapy","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — YAP epistasis via siRNA rescue, multiple functional readouts, single lab","pmids":["31807003"],"is_preprint":false},{"year":2019,"finding":"RASSF6 negatively regulates TRIM16 by promoting its ubiquitination-dependent degradation in esophageal squamous cell carcinoma cells, thereby activating cell cycle and EMT-associated pathways and promoting ESCC proliferation, migration, and invasion.","method":"IHC, OE/KD in ESCC cells, ubiquitination assay, in vivo xenograft and metastasis model","journal":"Journal of genetics and genomics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ubiquitination assay plus OE/KD functional phenotypes in vitro and in vivo, single lab","pmids":["31812473"],"is_preprint":false},{"year":2020,"finding":"UNC119A (a chaperone of myristoylated proteins) interacts with activated KRAS, promotes KRAS binding to RASSF6, and enhances the RASSF6-MDM2 interaction, driving p53-dependent apoptosis; UNC119A silencing promoted colony formation, migration, and invasion in KRAS-mutated cells.","method":"Immunoprecipitation, immunofluorescence, quantitative RT-PCR, RNAi gene silencing, soft-agar colony formation, migration and invasion assays","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, functional KD with multiple phenotypic readouts, single lab","pmids":["32554467"],"is_preprint":false},{"year":2021,"finding":"DNA damage triggers nuclear accumulation of RASSF6 via CDK9-mediated phosphorylation of BAF53; RASSF6 directly binds BAF53 (SWI/SNF complex component), the interaction is enhanced by CDK9, and RASSF6 augments BAF53-BAF60a interaction and their binding to p53, thereby promoting p53 target gene transcription and apoptosis; BAF53 or BAF60a silencing attenuates these effects.","method":"Nuclear/cytoplasmic fractionation, identification of nuclear localization and export signals, Co-immunoprecipitation, CDK9 inhibitor treatment, siRNA knockdown of BAF53/BAF60a, p53 target gene expression assays, apoptosis assays","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (fractionation, Co-IP, KD epistasis, gene expression), mechanistic chain fully dissected in single rigorous study","pmids":["34898277"],"is_preprint":false},{"year":2021,"finding":"Rassf6 knockout MEFs (generated by CRISPR/Cas9) show resistance to UV-induced apoptosis/cell cycle arrest and senescence, compromised p53-target gene expression, delayed DNA repair, and enhanced NF-κB signaling; KRAS active mutant promotes colony formation of Rassf6-null MEFs but not wild-type MEFs; Rassf6 knockout mice show enhanced skin inflammation upon DMBA treatment.","method":"CRISPR/Cas9 knockout mice, MEF cultures, UV treatment, RNA-seq, p53 target gene expression, soft-agar colony formation, in vivo DMBA-induced inflammation","journal":"Genes to cells : devoted to molecular & cellular mechanisms","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic KO model with multiple orthogonal in vitro and in vivo phenotypic readouts and RNA-seq pathway analysis","pmids":["34652874"],"is_preprint":false}],"current_model":"RASSF6 is a tumor suppressor that binds activated RAS (via its RA domain) and interacts with MST1/2 kinases to modulate the Hippo pathway; it stabilizes p53 by binding MDM2 and promoting its ubiquitination, cooperates with BAF53/BAF60a (SWI/SNF components) upon DNA damage to drive p53 target gene transcription, induces apoptosis and cell cycle arrest through pRb/p16/p14ARF axis in p53-negative contexts, activates JNK signaling to suppress Mcl-1 and other pro-survival factors, represses YAP downstream of the Hippo pathway, binds MAGI-1 via its PDZ-binding motif at cell junctions, and is antagonized by BCL-XL, with its nuclear localization regulated by CDK9-mediated BAF53 phosphorylation in response to DNA damage."},"narrative":{"mechanistic_narrative":"RASSF6 is a RAS-effector tumor suppressor that couples activated RAS to apoptosis and cell cycle arrest, with loss of RASSF6 enhancing tumorigenic and invasive phenotypes across multiple cancer types [PMID:17404571, PMID:34652874]. It operates at the interface of the Hippo and p53 pathways: RASSF6 binds the Hippo kinase MST2 and inhibits its activity, and upon MST2 activation is released to trigger WW45-dependent apoptosis, defining an apoptotic branch parallel to canonical Hippo signaling [PMID:19797269]; it also drives MST1/2 and LATS1 phosphorylation to repress the effector YAP, and YAP repression mediates RASSF6's downregulation of Bcl-xL and induction of mitochondrial apoptosis [PMID:29964010, PMID:31807003]. In parallel, RASSF6 binds MDM2 and promotes its ubiquitination, blocking MDM2-mediated p53 degradation to stabilize p53 and impose G1/S arrest, and is required for UV- and VP-16-induced apoptosis, DNA repair, and ploidy control [PMID:24003224, PMID:34652874]. Following DNA damage, CDK9-mediated phosphorylation of BAF53 drives nuclear accumulation of RASSF6, which binds BAF53 and bridges its association with BAF60a and p53 to potentiate p53 target-gene transcription [PMID:34898277]. In p53-negative contexts RASSF6 retains tumor-suppressor output through a pRb axis, enhancing pRb–phosphatase and pRb–E2F1 interactions and suppressing BMI1 to elevate P16INK4A and P14ARF [PMID:29891515]. RASSF6 additionally activates JNK signaling to suppress Mcl-1 and sensitize cells to apoptosis and sorafenib [PMID:24626183, PMID:29864454]. Its activity is antagonized by BCL-XL, which binds RASSF6 and blocks the RASSF6–MDM2 interaction [PMID:29193479], and its expression is repressed by oncogenic microRNAs including miR-181a-5p and miR-496 [PMID:28043911, PMID:31273789]. RASSF6 localizes to renal epithelial apical membranes and engages cell-junction scaffolds via a C-terminal PDZ-binding motif that binds the PDZ domains of MAGI-1 [PMID:22577168].","teleology":[{"year":2007,"claim":"Established RASSF6 as a RAS-association-domain effector that links activated RAS to apoptosis, raising the question of whether it acts as a tumor suppressor.","evidence":"Exogenous expression and suppression in lung tumor cell lines with cell death assays","pmids":["17404571"],"confidence":"Medium","gaps":["Downstream apoptotic effectors not defined","RAS-binding to growth-suppression mechanism not connected","Single cell-line context"]},{"year":2009,"claim":"Placed RASSF6 within Hippo signaling by showing it inhibits MST2 and is released upon MST2 activation to trigger WW45-dependent apoptosis, defining a parallel apoptotic branch.","evidence":"Reciprocal Co-IP, kinase activity assays, and epistasis with WW45","pmids":["19797269"],"confidence":"High","gaps":["How RAS binding releases RASSF6 from MST2 not resolved","Connection to downstream YAP not yet tested","Endogenous stoichiometry unclear"]},{"year":2012,"claim":"Identified a junctional scaffolding role through PDZ-binding-motif-dependent MAGI-1 binding and defined RASSF6's epithelial localization.","evidence":"Yeast two-hybrid, Co-IP from rat liver, kidney immunolocalization, HK-2 apoptosis assays","pmids":["22577168"],"confidence":"Medium","gaps":["Functional consequence of MAGI-1 binding not dissected","Link between junctional localization and apoptotic signaling unclear"]},{"year":2013,"claim":"Connected RASSF6 to the p53 axis by showing it binds MDM2, promotes MDM2 ubiquitination, and stabilizes p53 to enforce arrest and DNA-damage responses.","evidence":"Co-IP, ubiquitination assays, siRNA knockdown, cell cycle and p53 target gene analysis","pmids":["24003224"],"confidence":"High","gaps":["Whether RASSF6 acts as a direct E3 cofactor not determined","Structural basis of MDM2 binding unknown"]},{"year":2014,"claim":"Defined a RASSF6-JNK arm driving apoptosis and p21-dependent G1 arrest, broadening the tumor-suppressor output beyond p53.","evidence":"Ectopic re-expression/knockdown, JNK inhibitor, cell cycle analysis, xenografts in RCC","pmids":["24626183"],"confidence":"Medium","gaps":["How RASSF6 activates JNK upstream not defined","Crosstalk with Hippo/p53 arms not mapped"]},{"year":2014,"claim":"Showed RASSF6 re-expression suppresses MAPK/AKT signaling and invasion in BRAF-mutant melanoma, increasing mutant B-Raf–MST1 association.","evidence":"Collagen invasion assay, MAPK/AKT western blot, B-Raf/MST1 Co-IP","pmids":["25482183"],"confidence":"Medium","gaps":["Direct vs indirect MAPK suppression unresolved","Mechanism of B-Raf–MST1 bridging unclear"]},{"year":2016,"claim":"Established RASSF6 as a target of oncogenic microRNAs, with miR-181a-5p directly repressing it to activate MAPK and drive gastric cancer progression.","evidence":"Luciferase 3'UTR reporter, miRNA mimic/inhibitor with RASSF6 rescue, in vivo assays","pmids":["28043911"],"confidence":"Medium","gaps":["Whether miRNA repression accounts for tumor RASSF6 loss generally not established"]},{"year":2018,"claim":"Revealed a p53-independent tumor-suppressor route via pRb: RASSF6 promotes pRb dephosphorylation, suppresses BMI1, and elevates P16INK4A/P14ARF.","evidence":"Co-IP, western blot, siRNA, flow cytometry in p53-negative cells","pmids":["29891515"],"confidence":"High","gaps":["How RASSF6 enhances pRb-phosphatase interaction mechanistically unknown","Relationship between pRb and Hippo arms unclear"]},{"year":2018,"claim":"Linked RASSF6-JNK signaling to Mcl-1 suppression and sorafenib sensitivity, establishing a RASSF6-JNK-Mcl-1 axis.","evidence":"Stable overexpression, JNK inhibitor, Mcl-1 assays, in vitro/in vivo sorafenib resistance","pmids":["29864454"],"confidence":"Medium","gaps":["Direct mechanism of Mcl-1 downregulation not defined"]},{"year":2018,"claim":"Positioned YAP downstream of RASSF6, showing RASSF6 activates MST1/2-LATS1 to repress YAP and that YAP repression mediates Bcl-xL downregulation and mitochondrial apoptosis.","evidence":"Overexpression/depletion, YAP rescue epistasis, western blot, JC-1 staining in breast cancer cells","pmids":["29964010"],"confidence":"Medium","gaps":["Reconciliation with MST2-inhibitory role from 2009 not addressed","Direct LATS1 activation mechanism unclear"]},{"year":2019,"claim":"Confirmed YAP epistasis in a second cancer type, with YAP silencing abolishing RASSF6 effects on Bcl-xL and apoptosis.","evidence":"OE/siRNA, YAP rescue, JC-1, Annexin V/PI in bladder cancer cells","pmids":["31807003"],"confidence":"Medium","gaps":["Context dependence of RASSF6-YAP regulation across tissues unmapped"]},{"year":2019,"claim":"Extended miRNA-mediated silencing to miR-496, which directly represses RASSF6 to activate Wnt signaling in colorectal cancer.","evidence":"Luciferase reporter, miR-496 mimic/inhibitor, RASSF6 siRNA rescue, migration/EMT assays","pmids":["31273789"],"confidence":"Medium","gaps":["Link between RASSF6 loss and Wnt activation not mechanistically dissected"]},{"year":2019,"claim":"Reported a context where RASSF6 promotes tumor progression by driving ubiquitination-dependent degradation of TRIM16 in esophageal carcinoma.","evidence":"IHC, OE/KD, ubiquitination assay, xenograft and metastasis models","pmids":["31812473"],"confidence":"Medium","gaps":["Apparent oncogenic role conflicts with tumor-suppressor findings elsewhere","Mechanism of TRIM16 ubiquitin-targeting unclear"]},{"year":2020,"claim":"Showed UNC119A bridges activated KRAS to RASSF6 and enhances the RASSF6-MDM2 interaction to drive p53-dependent apoptosis, connecting RAS sensing to p53 stabilization.","evidence":"Co-IP, immunofluorescence, RNAi, soft-agar, migration/invasion assays in KRAS-mutant cells","pmids":["32554467"],"confidence":"Medium","gaps":["Structural basis of the KRAS-UNC119A-RASSF6 assembly unknown"]},{"year":2021,"claim":"Dissected the DNA-damage-triggered nuclear program, showing CDK9-phosphorylated BAF53 drives RASSF6 nuclear accumulation and that RASSF6 bridges BAF53-BAF60a-p53 to promote p53 target transcription.","evidence":"Fractionation, NLS/NES mapping, Co-IP, CDK9 inhibitor, BAF53/BAF60a siRNA, p53 target and apoptosis assays","pmids":["34898277"],"confidence":"High","gaps":["Direct chromatin recruitment to p53 target promoters not shown","Generality across damage types unclear"]},{"year":2021,"claim":"Provided in vivo genetic confirmation that Rassf6 is required for UV-induced p53 responses, DNA repair, and senescence and restrains KRAS-driven transformation and skin inflammation.","evidence":"CRISPR/Cas9 knockout mice and MEFs, RNA-seq, soft-agar, DMBA-induced inflammation","pmids":["34652874"],"confidence":"High","gaps":["Tissue-specific contributions not separated","Enhanced NF-kB signaling mechanism not defined"]},{"year":null,"claim":"How RASSF6 reconciles its dual reported roles—tumor suppressor (MST2/p53/pRb/JNK) versus context-specific oncogenic TRIM16 degradation—and what determines its choice between Hippo, p53, and JNK outputs remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural model of RASSF6 multi-partner assemblies","Determinants of context-dependent output unknown","Direct biochemical activity (E3 cofactor vs scaffold) undefined"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[3,17,16]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[1,3,9]}],"localization":[{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[17]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[17]},{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[2]}],"pathway":[],"complexes":[],"partners":["MST2","MDM2","MAGI1","BCL2L1","BAF53","SMARCD1","KRAS","UNC119"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q6ZTQ3","full_name":"Ras association domain-containing protein 6","aliases":[],"length_aa":369,"mass_kda":43.4,"function":"Involved in the induction of apoptosis, through both caspase-dependent and caspase-independent pathways. May act as a Ras effector protein. May suppress the serum-induced basal levels of NF-kappa-B (By similarity)","subcellular_location":"","url":"https://www.uniprot.org/uniprotkb/Q6ZTQ3/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/RASSF6","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"STK4","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/RASSF6","total_profiled":1310},"omim":[{"mim_id":"619504","title":"CHOPRA-AMIEL-GORDON SYNDROME; CAGS","url":"https://www.omim.org/entry/619504"},{"mim_id":"615929","title":"ANKYRIN REPEAT DOMAIN-CONTAINING PROTEIN 17; ANKRD17","url":"https://www.omim.org/entry/615929"},{"mim_id":"614713","title":"RAS ASSOCIATION DOMAIN FAMILY, MEMBER 10; RASSF10","url":"https://www.omim.org/entry/614713"},{"mim_id":"612620","title":"RAS ASSOCIATION DOMAIN FAMILY, MEMBER 6; RASSF6","url":"https://www.omim.org/entry/612620"},{"mim_id":"162332","title":"TACHYKININ RECEPTOR 3; TACR3","url":"https://www.omim.org/entry/162332"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Nucleoplasm","reliability":"Approved"},{"location":"Golgi apparatus","reliability":"Approved"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"lymphoid tissue","ntpm":28.5},{"tissue":"stomach 1","ntpm":33.0}],"url":"https://www.proteinatlas.org/search/RASSF6"},"hgnc":{"alias_symbol":[],"prev_symbol":[]},"alphafold":{"accession":"Q6ZTQ3","domains":[{"cath_id":"3.10.20.90","chopping":"44-105_223-312","consensus_level":"high","plddt":89.7161,"start":44,"end":312},{"cath_id":"1.20.5","chopping":"320-369","consensus_level":"medium","plddt":88.4284,"start":320,"end":369}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q6ZTQ3","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q6ZTQ3-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q6ZTQ3-F1-predicted_aligned_error_v6.png","plddt_mean":70.06},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=RASSF6","jax_strain_url":"https://www.jax.org/strain/search?query=RASSF6"},"sequence":{"accession":"Q6ZTQ3","fasta_url":"https://rest.uniprot.org/uniprotkb/Q6ZTQ3.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q6ZTQ3/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q6ZTQ3"}},"corpus_meta":[{"pmid":"17404571","id":"PMC_17404571","title":"RASSF6 is a novel member of the RASSF family of tumor suppressors.","date":"2007","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/17404571","citation_count":114,"is_preprint":false},{"pmid":"19570220","id":"PMC_19570220","title":"The novel RASSF6 and RASSF10 candidate tumour suppressor genes are frequently epigenetically inactivated in childhood leukaemias.","date":"2009","source":"Molecular cancer","url":"https://pubmed.ncbi.nlm.nih.gov/19570220","citation_count":91,"is_preprint":false},{"pmid":"28043911","id":"PMC_28043911","title":"miR-181a-5p promotes the progression of gastric cancer via RASSF6-mediated MAPK signalling activation.","date":"2016","source":"Cancer letters","url":"https://pubmed.ncbi.nlm.nih.gov/28043911","citation_count":90,"is_preprint":false},{"pmid":"19797269","id":"PMC_19797269","title":"Hippo pathway-dependent and -independent roles of RASSF6.","date":"2009","source":"Science signaling","url":"https://pubmed.ncbi.nlm.nih.gov/19797269","citation_count":84,"is_preprint":false},{"pmid":"22695170","id":"PMC_22695170","title":"The RASSF gene family members RASSF5, RASSF6 and RASSF7 show frequent DNA methylation in neuroblastoma.","date":"2012","source":"Molecular cancer","url":"https://pubmed.ncbi.nlm.nih.gov/22695170","citation_count":64,"is_preprint":false},{"pmid":"25482183","id":"PMC_25482183","title":"RASSF6 exhibits promoter hypermethylation in metastatic melanoma and inhibits invasion in melanoma cells.","date":"2014","source":"Epigenetics","url":"https://pubmed.ncbi.nlm.nih.gov/25482183","citation_count":37,"is_preprint":false},{"pmid":"24003224","id":"PMC_24003224","title":"The RASSF6 tumor suppressor protein regulates apoptosis and the cell cycle via MDM2 protein and p53 protein.","date":"2013","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/24003224","citation_count":35,"is_preprint":false},{"pmid":"31273789","id":"PMC_31273789","title":"MiR-496 promotes migration and epithelial-mesenchymal transition by targeting RASSF6 in colorectal cancer.","date":"2019","source":"Journal of cellular physiology","url":"https://pubmed.ncbi.nlm.nih.gov/31273789","citation_count":30,"is_preprint":false},{"pmid":"26690221","id":"PMC_26690221","title":"RASSF6; the Putative Tumor Suppressor of the RASSF Family.","date":"2015","source":"Cancers","url":"https://pubmed.ncbi.nlm.nih.gov/26690221","citation_count":19,"is_preprint":false},{"pmid":"27009808","id":"PMC_27009808","title":"Decreased level of RASSF6 in sporadic colorectal cancer and its anti-tumor effects both in vitro and in vivo.","date":"2016","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/27009808","citation_count":19,"is_preprint":false},{"pmid":"24626183","id":"PMC_24626183","title":"RASSF6 promotes p21(Cip1/Waf1)-dependent cell cycle arrest and apoptosis through activation of the JNK/SAPK pathway in clear cell renal cell carcinoma.","date":"2014","source":"Cell cycle (Georgetown, Tex.)","url":"https://pubmed.ncbi.nlm.nih.gov/24626183","citation_count":19,"is_preprint":false},{"pmid":"29891515","id":"PMC_29891515","title":"The RASSF6 Tumor Suppressor Protein Regulates Apoptosis and Cell Cycle Progression via Retinoblastoma Protein.","date":"2018","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/29891515","citation_count":18,"is_preprint":false},{"pmid":"22577168","id":"PMC_22577168","title":"Expression of RASSF6 in kidney and the implication of RASSF6 and the Hippo pathway in the sorbitol-induced apoptosis in renal proximal tubular epithelial cells.","date":"2012","source":"Journal of biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/22577168","citation_count":18,"is_preprint":false},{"pmid":"34151476","id":"PMC_34151476","title":"Down-regulation of circITCH promotes osteosarcoma development and resistance to doxorubicin via the miR-524/RASSF6 axis.","date":"2021","source":"The journal of gene medicine","url":"https://pubmed.ncbi.nlm.nih.gov/34151476","citation_count":18,"is_preprint":false},{"pmid":"29964010","id":"PMC_29964010","title":"Downregulation of RASSF6 promotes breast cancer growth and chemoresistance through regulation of Hippo signaling.","date":"2018","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/29964010","citation_count":17,"is_preprint":false},{"pmid":"23626755","id":"PMC_23626755","title":"RASSF6 expression in adipocytes is down-regulated by interaction with macrophages.","date":"2013","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/23626755","citation_count":14,"is_preprint":false},{"pmid":"25028967","id":"PMC_25028967","title":"Downregulation of Ras association domain family member 6 (RASSF6) underlies the treatment resistance of highly metastatic nasopharyngeal carcinoma cells.","date":"2014","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/25028967","citation_count":14,"is_preprint":false},{"pmid":"29864454","id":"PMC_29864454","title":"RASSF6-mediated inhibition of Mcl-1 through JNK activation improves the anti-tumor effects of sorafenib in renal cell carcinoma.","date":"2018","source":"Cancer letters","url":"https://pubmed.ncbi.nlm.nih.gov/29864454","citation_count":13,"is_preprint":false},{"pmid":"31807003","id":"PMC_31807003","title":"RASSF6 Is Downregulated In Human Bladder Cancers And Regulates Doxorubicin Sensitivity And Mitochondrial Membrane Potential Via The Hippo Signaling Pathway.","date":"2019","source":"OncoTargets and therapy","url":"https://pubmed.ncbi.nlm.nih.gov/31807003","citation_count":12,"is_preprint":false},{"pmid":"33416181","id":"PMC_33416181","title":"lncRNA TUSC7 inhibits osteosarcoma progression through the miR‑181a/RASSF6 axis.","date":"2020","source":"International journal of molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/33416181","citation_count":11,"is_preprint":false},{"pmid":"31812473","id":"PMC_31812473","title":"RASSF6-TRIM16 axis promotes cell proliferation, migration and invasion in esophageal squamous cell carcinoma.","date":"2019","source":"Journal of genetics and genomics = Yi chuan xue bao","url":"https://pubmed.ncbi.nlm.nih.gov/31812473","citation_count":10,"is_preprint":false},{"pmid":"29193479","id":"PMC_29193479","title":"BCL-XL binds and antagonizes RASSF6 tumor suppressor to suppress p53 expression.","date":"2017","source":"Genes to cells : devoted to molecular & cellular mechanisms","url":"https://pubmed.ncbi.nlm.nih.gov/29193479","citation_count":9,"is_preprint":false},{"pmid":"31486880","id":"PMC_31486880","title":"Residual methylation of tumor suppressor gene promoters, RASSF6 and RASSF10, as novel biomarkers for minimal residual disease detection in adult acute lymphoblastic leukemia.","date":"2019","source":"Annals of hematology","url":"https://pubmed.ncbi.nlm.nih.gov/31486880","citation_count":9,"is_preprint":false},{"pmid":"27983932","id":"PMC_27983932","title":"Overexpression of RAS-Association Domain Family 6 (RASSF6) Inhibits Proliferation and Tumorigenesis in Hepatocellular Carcinoma Cells.","date":"2016","source":"Oncology research","url":"https://pubmed.ncbi.nlm.nih.gov/27983932","citation_count":8,"is_preprint":false},{"pmid":"34898277","id":"PMC_34898277","title":"DNA Damage Triggers the Nuclear Accumulation of RASSF6 Tumor Suppressor Protein via CDK9 and BAF53 To Regulate p53 Target Gene Transcription.","date":"2021","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/34898277","citation_count":4,"is_preprint":false},{"pmid":"32554467","id":"PMC_32554467","title":"The RAS-interacting chaperone UNC119 drives the RASSF6-MDM2-p53 axis and antagonizes RAS-mediated malignant transformation.","date":"2020","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/32554467","citation_count":4,"is_preprint":false},{"pmid":"35048636","id":"PMC_35048636","title":"Kindlin-3 and RASSF6 are probable biomarkers for predicting metastasis in cutaneous melanoma.","date":"2021","source":"Polish journal of pathology : official journal of the Polish Society of Pathologists","url":"https://pubmed.ncbi.nlm.nih.gov/35048636","citation_count":3,"is_preprint":false},{"pmid":"40067291","id":"PMC_40067291","title":"Induction of miR-224 by Reactive Oxygen Species Regulates RASSF6 and Thus Modulates Malignant Behaviors and Chemosensitivity in Esophageal Squamous Cell Carcinoma.","date":"2025","source":"Journal of biochemical and molecular toxicology","url":"https://pubmed.ncbi.nlm.nih.gov/40067291","citation_count":2,"is_preprint":false},{"pmid":"41630709","id":"PMC_41630709","title":"Apoptotic role of RASSF6 in regulating PRV-infected porcine alveolar macrophages.","date":"2025","source":"Open veterinary journal","url":"https://pubmed.ncbi.nlm.nih.gov/41630709","citation_count":0,"is_preprint":false},{"pmid":"34652874","id":"PMC_34652874","title":"Characterization of mouse embryonic fibroblasts derived from Rassf6 knockout mice shows the implication of Rassf6 in the regulation of NF-κB signaling.","date":"2021","source":"Genes to cells : devoted to molecular & cellular mechanisms","url":"https://pubmed.ncbi.nlm.nih.gov/34652874","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":17069,"output_tokens":4573,"usd":0.059901,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":12554,"output_tokens":4750,"usd":0.09076,"stage2_stop_reason":"end_turn"},"total_usd":0.150661,"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\": 2007,\n      \"finding\": \"RASSF6 possesses a Ras Association domain and binds activated Ras, promoting apoptosis and synergizing with activated K-Ras to induce cell death; suppression of RASSF6 enhanced the tumorigenic phenotype of a human lung tumor cell line.\",\n      \"method\": \"Exogenous expression, suppression experiments in tumor cell lines, cell death assays\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional KD/OE with defined phenotype and Ras-binding characterization in single lab\",\n      \"pmids\": [\"17404571\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"RASSF6 binds MST2 (mammalian Hippo kinase homolog) and inhibits MST2 kinase activity, antagonizing Hippo signaling; upon MST2 activation, RASSF6 is released and causes apoptosis in a WW45 (mammalian Salvador homolog)-dependent manner, defining a RASSF6-mediated apoptotic pathway parallel to the canonical Hippo pathway.\",\n      \"method\": \"Co-immunoprecipitation, kinase activity assays, epistasis analysis with WW45\",\n      \"journal\": \"Science signaling\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal Co-IP, kinase assays, and genetic epistasis in single lab with multiple orthogonal methods\",\n      \"pmids\": [\"19797269\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"RASSF6 binds PDZ domains of MAGI-1 through its C-terminal PDZ-binding motif and co-immunoprecipitates with MAGI-1 from rat liver; RASSF6 is localized in normal kidney glomerulus and on apical membranes of renal proximal tubular epithelial cells, and is involved in sorbitol-induced apoptosis together with the Hippo pathway in HK-2 cells.\",\n      \"method\": \"Yeast two-hybrid screening (original identification), co-immunoprecipitation from rat liver, immunolocalization in kidney, functional apoptosis assays in HK-2 cells\",\n      \"journal\": \"Journal of biochemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP from tissue, localization, and functional assay in single lab with multiple orthogonal methods\",\n      \"pmids\": [\"22577168\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"RASSF6 binds MDM2 and facilitates its ubiquitination, thereby blocking MDM2-mediated p53 degradation and stabilizing p53; RASSF6 overexpression induces G1/S arrest in p53-positive cells, and its depletion prevents UV- and VP-16-induced apoptosis and G1/S arrest, delays DNA repair, and increases polyploid cells.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assays, siRNA knockdown, cell cycle analysis, p53 target gene expression\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, ubiquitination assay, KD with multiple defined phenotypic readouts in single lab\",\n      \"pmids\": [\"24003224\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"RASSF6 mRNA in adipocytes is downregulated by conditioned medium from activated macrophages, and decreased RASSF6 expression upregulates CD44 and HMGA2 mRNA expression in adipocytes.\",\n      \"method\": \"Gene expression profiling in 3T3-L1 adipocytes co-cultured with RAW264.7 macrophages, in vivo correlation with macrophage number\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — expression-level findings with downstream gene targets noted, single lab, no direct mechanistic pathway dissection\",\n      \"pmids\": [\"23626755\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"RASSF6 activates JNK signaling to promote apoptosis and p21(Cip1/Waf1)-dependent G1 cell cycle arrest in clear cell renal cell carcinoma cells; JNK inhibitor SP600125 blocked RASSF6-mediated apoptosis sensitization.\",\n      \"method\": \"Ectopic re-expression and siRNA knockdown, JNK inhibitor treatment, cell cycle analysis, tumor xenograft models\",\n      \"journal\": \"Cell cycle (Georgetown, Tex.)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — KD/OE with pharmacological inhibition of JNK and multiple phenotypic readouts, single lab\",\n      \"pmids\": [\"24626183\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"RASSF6 re-expression in metastatic melanoma cells with B-Raf(V600E) inhibited invasion through collagen, suppressed MAPK pathway activation and AKT, and increased association of mutant B-Raf with MST1.\",\n      \"method\": \"Invasion assay through collagen, western blot for MAPK/AKT, co-immunoprecipitation of B-Raf and MST1\",\n      \"journal\": \"Epigenetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, functional invasion assay, and signaling readouts in single lab\",\n      \"pmids\": [\"25482183\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"miR-181a-5p directly represses RASSF6 expression and thereby activates the MAPK signaling pathway, promoting gastric cancer cell proliferation, invasion, and EMT; overexpression or silencing of RASSF6 partially reverses the oncogenic effects of miR-181a-5p.\",\n      \"method\": \"Luciferase reporter assay for direct miR-181a-5p binding to RASSF6 3'UTR, miRNA mimic/inhibitor with rescue by RASSF6 OE/KD, in vitro and in vivo assays\",\n      \"journal\": \"Cancer letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — luciferase reporter (direct binding validation), rescue experiments, in vivo confirmation, single lab\",\n      \"pmids\": [\"28043911\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"RASSF6 overexpression in HCC cells suppresses FAK phosphorylation and decreases MMP-2 and MMP-9 expression, inhibiting invasion and migration.\",\n      \"method\": \"Overexpression in HepG2 cells, western blot for p-FAK, MMP-2, MMP-9, invasion assays, xenograft model\",\n      \"journal\": \"Oncology research\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, OE only with western blot readouts, no direct mechanistic dissection of FAK regulation\",\n      \"pmids\": [\"27983932\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"BCL-XL directly interacts with RASSF6, inhibits the interaction between RASSF6 and MDM2, and suppresses p53 expression, thereby antagonizing RASSF6-mediated apoptosis.\",\n      \"method\": \"Co-immunoprecipitation, p53 expression assays, apoptosis assays with BCL-XL manipulation\",\n      \"journal\": \"Genes to cells : devoted to molecular & cellular mechanisms\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP showing direct interaction and functional consequence on p53/apoptosis, single lab\",\n      \"pmids\": [\"29193479\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"RASSF6 mediates cell cycle arrest and apoptosis via pRb independent of p53: RASSF6 enhances the interaction between pRb and protein phosphatase, suppresses BMI1 to increase P16INK4A and P14ARF expression, increases unphosphorylated pRb, augments pRb-E2F1 interaction, and induces TP73 target genes in a p53-negative background.\",\n      \"method\": \"Co-immunoprecipitation, western blot, siRNA knockdown, flow cytometry cell cycle analysis in p53-negative cells\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (Co-IP, gene expression, cell cycle), p53-negative background controls, multiple pathway nodes tested in single rigorous study\",\n      \"pmids\": [\"29891515\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"RASSF6 activates the JNK signaling pathway, which contributes to Mcl-1 inhibition; pharmacological JNK pathway suppression partially restores Mcl-1 expression and sorafenib resistance, establishing a RASSF6-JNK-Mcl-1 axis.\",\n      \"method\": \"Stable RASSF6 overexpression in RCC cell lines, JNK inhibitor treatment, Mcl-1 expression assays, in vitro and in vivo sorafenib resistance assays\",\n      \"journal\": \"Cancer letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — stable OE, pharmacological rescue, in vitro and in vivo validation, single lab\",\n      \"pmids\": [\"29864454\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"RASSF6 activates Hippo signaling by upregulating MST1/2 and LATS1 phosphorylation, represses YAP, and YAP repression mediates RASSF6's effects on Bcl-xL, apoptosis (cleaved caspase-3, cytochrome c), and mitochondrial membrane potential in breast cancer cells.\",\n      \"method\": \"Overexpression/depletion of RASSF6, YAP rescue experiments, western blot, JC-1 staining, flow cytometry\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — YAP rescue epistasis, multiple pathway markers, single lab\",\n      \"pmids\": [\"29964010\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"miR-496 directly binds RASSF6 and represses its expression, activating Wnt signaling to promote CRC cell migration and EMT; this effect was validated by luciferase reporter assay and RASSF6 siRNA rescue experiments.\",\n      \"method\": \"Luciferase reporter assay, miR-496 mimic/inhibitor, RASSF6 siRNA rescue, migration and EMT assays\",\n      \"journal\": \"Journal of cellular physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct binding validated by luciferase reporter, functional rescue, single lab\",\n      \"pmids\": [\"31273789\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"RASSF6 overexpression in bladder cancer cells downregulates YAP (Hippo pathway effector) and Bcl-xL; YAP silencing abolishes the effects of RASSF6 on Bcl-xL expression, doxorubicin-induced apoptosis, and mitochondrial membrane potential, placing YAP downstream of RASSF6.\",\n      \"method\": \"Overexpression/siRNA knockdown, YAP rescue/epistasis, JC-1 staining, western blot, AnnexinV/PI apoptosis assay\",\n      \"journal\": \"OncoTargets and therapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — YAP epistasis via siRNA rescue, multiple functional readouts, single lab\",\n      \"pmids\": [\"31807003\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"RASSF6 negatively regulates TRIM16 by promoting its ubiquitination-dependent degradation in esophageal squamous cell carcinoma cells, thereby activating cell cycle and EMT-associated pathways and promoting ESCC proliferation, migration, and invasion.\",\n      \"method\": \"IHC, OE/KD in ESCC cells, ubiquitination assay, in vivo xenograft and metastasis model\",\n      \"journal\": \"Journal of genetics and genomics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ubiquitination assay plus OE/KD functional phenotypes in vitro and in vivo, single lab\",\n      \"pmids\": [\"31812473\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"UNC119A (a chaperone of myristoylated proteins) interacts with activated KRAS, promotes KRAS binding to RASSF6, and enhances the RASSF6-MDM2 interaction, driving p53-dependent apoptosis; UNC119A silencing promoted colony formation, migration, and invasion in KRAS-mutated cells.\",\n      \"method\": \"Immunoprecipitation, immunofluorescence, quantitative RT-PCR, RNAi gene silencing, soft-agar colony formation, migration and invasion assays\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, functional KD with multiple phenotypic readouts, single lab\",\n      \"pmids\": [\"32554467\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"DNA damage triggers nuclear accumulation of RASSF6 via CDK9-mediated phosphorylation of BAF53; RASSF6 directly binds BAF53 (SWI/SNF complex component), the interaction is enhanced by CDK9, and RASSF6 augments BAF53-BAF60a interaction and their binding to p53, thereby promoting p53 target gene transcription and apoptosis; BAF53 or BAF60a silencing attenuates these effects.\",\n      \"method\": \"Nuclear/cytoplasmic fractionation, identification of nuclear localization and export signals, Co-immunoprecipitation, CDK9 inhibitor treatment, siRNA knockdown of BAF53/BAF60a, p53 target gene expression assays, apoptosis assays\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (fractionation, Co-IP, KD epistasis, gene expression), mechanistic chain fully dissected in single rigorous study\",\n      \"pmids\": [\"34898277\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Rassf6 knockout MEFs (generated by CRISPR/Cas9) show resistance to UV-induced apoptosis/cell cycle arrest and senescence, compromised p53-target gene expression, delayed DNA repair, and enhanced NF-κB signaling; KRAS active mutant promotes colony formation of Rassf6-null MEFs but not wild-type MEFs; Rassf6 knockout mice show enhanced skin inflammation upon DMBA treatment.\",\n      \"method\": \"CRISPR/Cas9 knockout mice, MEF cultures, UV treatment, RNA-seq, p53 target gene expression, soft-agar colony formation, in vivo DMBA-induced inflammation\",\n      \"journal\": \"Genes to cells : devoted to molecular & cellular mechanisms\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic KO model with multiple orthogonal in vitro and in vivo phenotypic readouts and RNA-seq pathway analysis\",\n      \"pmids\": [\"34652874\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"RASSF6 is a tumor suppressor that binds activated RAS (via its RA domain) and interacts with MST1/2 kinases to modulate the Hippo pathway; it stabilizes p53 by binding MDM2 and promoting its ubiquitination, cooperates with BAF53/BAF60a (SWI/SNF components) upon DNA damage to drive p53 target gene transcription, induces apoptosis and cell cycle arrest through pRb/p16/p14ARF axis in p53-negative contexts, activates JNK signaling to suppress Mcl-1 and other pro-survival factors, represses YAP downstream of the Hippo pathway, binds MAGI-1 via its PDZ-binding motif at cell junctions, and is antagonized by BCL-XL, with its nuclear localization regulated by CDK9-mediated BAF53 phosphorylation in response to DNA damage.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"RASSF6 is a RAS-effector tumor suppressor that couples activated RAS to apoptosis and cell cycle arrest, with loss of RASSF6 enhancing tumorigenic and invasive phenotypes across multiple cancer types [#0, #18]. It operates at the interface of the Hippo and p53 pathways: RASSF6 binds the Hippo kinase MST2 and inhibits its activity, and upon MST2 activation is released to trigger WW45-dependent apoptosis, defining an apoptotic branch parallel to canonical Hippo signaling [#1]; it also drives MST1/2 and LATS1 phosphorylation to repress the effector YAP, and YAP repression mediates RASSF6's downregulation of Bcl-xL and induction of mitochondrial apoptosis [#12, #14]. In parallel, RASSF6 binds MDM2 and promotes its ubiquitination, blocking MDM2-mediated p53 degradation to stabilize p53 and impose G1/S arrest, and is required for UV- and VP-16-induced apoptosis, DNA repair, and ploidy control [#3, #18]. Following DNA damage, CDK9-mediated phosphorylation of BAF53 drives nuclear accumulation of RASSF6, which binds BAF53 and bridges its association with BAF60a and p53 to potentiate p53 target-gene transcription [#17]. In p53-negative contexts RASSF6 retains tumor-suppressor output through a pRb axis, enhancing pRb–phosphatase and pRb–E2F1 interactions and suppressing BMI1 to elevate P16INK4A and P14ARF [#10]. RASSF6 additionally activates JNK signaling to suppress Mcl-1 and sensitize cells to apoptosis and sorafenib [#5, #11]. Its activity is antagonized by BCL-XL, which binds RASSF6 and blocks the RASSF6–MDM2 interaction [#9], and its expression is repressed by oncogenic microRNAs including miR-181a-5p and miR-496 [#7, #13]. RASSF6 localizes to renal epithelial apical membranes and engages cell-junction scaffolds via a C-terminal PDZ-binding motif that binds the PDZ domains of MAGI-1 [#2].\",\n  \"teleology\": [\n    {\n      \"year\": 2007,\n      \"claim\": \"Established RASSF6 as a RAS-association-domain effector that links activated RAS to apoptosis, raising the question of whether it acts as a tumor suppressor.\",\n      \"evidence\": \"Exogenous expression and suppression in lung tumor cell lines with cell death assays\",\n      \"pmids\": [\"17404571\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Downstream apoptotic effectors not defined\", \"RAS-binding to growth-suppression mechanism not connected\", \"Single cell-line context\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Placed RASSF6 within Hippo signaling by showing it inhibits MST2 and is released upon MST2 activation to trigger WW45-dependent apoptosis, defining a parallel apoptotic branch.\",\n      \"evidence\": \"Reciprocal Co-IP, kinase activity assays, and epistasis with WW45\",\n      \"pmids\": [\"19797269\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How RAS binding releases RASSF6 from MST2 not resolved\", \"Connection to downstream YAP not yet tested\", \"Endogenous stoichiometry unclear\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Identified a junctional scaffolding role through PDZ-binding-motif-dependent MAGI-1 binding and defined RASSF6's epithelial localization.\",\n      \"evidence\": \"Yeast two-hybrid, Co-IP from rat liver, kidney immunolocalization, HK-2 apoptosis assays\",\n      \"pmids\": [\"22577168\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional consequence of MAGI-1 binding not dissected\", \"Link between junctional localization and apoptotic signaling unclear\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Connected RASSF6 to the p53 axis by showing it binds MDM2, promotes MDM2 ubiquitination, and stabilizes p53 to enforce arrest and DNA-damage responses.\",\n      \"evidence\": \"Co-IP, ubiquitination assays, siRNA knockdown, cell cycle and p53 target gene analysis\",\n      \"pmids\": [\"24003224\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether RASSF6 acts as a direct E3 cofactor not determined\", \"Structural basis of MDM2 binding unknown\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Defined a RASSF6-JNK arm driving apoptosis and p21-dependent G1 arrest, broadening the tumor-suppressor output beyond p53.\",\n      \"evidence\": \"Ectopic re-expression/knockdown, JNK inhibitor, cell cycle analysis, xenografts in RCC\",\n      \"pmids\": [\"24626183\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"How RASSF6 activates JNK upstream not defined\", \"Crosstalk with Hippo/p53 arms not mapped\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Showed RASSF6 re-expression suppresses MAPK/AKT signaling and invasion in BRAF-mutant melanoma, increasing mutant B-Raf–MST1 association.\",\n      \"evidence\": \"Collagen invasion assay, MAPK/AKT western blot, B-Raf/MST1 Co-IP\",\n      \"pmids\": [\"25482183\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct vs indirect MAPK suppression unresolved\", \"Mechanism of B-Raf–MST1 bridging unclear\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Established RASSF6 as a target of oncogenic microRNAs, with miR-181a-5p directly repressing it to activate MAPK and drive gastric cancer progression.\",\n      \"evidence\": \"Luciferase 3'UTR reporter, miRNA mimic/inhibitor with RASSF6 rescue, in vivo assays\",\n      \"pmids\": [\"28043911\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether miRNA repression accounts for tumor RASSF6 loss generally not established\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Revealed a p53-independent tumor-suppressor route via pRb: RASSF6 promotes pRb dephosphorylation, suppresses BMI1, and elevates P16INK4A/P14ARF.\",\n      \"evidence\": \"Co-IP, western blot, siRNA, flow cytometry in p53-negative cells\",\n      \"pmids\": [\"29891515\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How RASSF6 enhances pRb-phosphatase interaction mechanistically unknown\", \"Relationship between pRb and Hippo arms unclear\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Linked RASSF6-JNK signaling to Mcl-1 suppression and sorafenib sensitivity, establishing a RASSF6-JNK-Mcl-1 axis.\",\n      \"evidence\": \"Stable overexpression, JNK inhibitor, Mcl-1 assays, in vitro/in vivo sorafenib resistance\",\n      \"pmids\": [\"29864454\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct mechanism of Mcl-1 downregulation not defined\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Positioned YAP downstream of RASSF6, showing RASSF6 activates MST1/2-LATS1 to repress YAP and that YAP repression mediates Bcl-xL downregulation and mitochondrial apoptosis.\",\n      \"evidence\": \"Overexpression/depletion, YAP rescue epistasis, western blot, JC-1 staining in breast cancer cells\",\n      \"pmids\": [\"29964010\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Reconciliation with MST2-inhibitory role from 2009 not addressed\", \"Direct LATS1 activation mechanism unclear\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Confirmed YAP epistasis in a second cancer type, with YAP silencing abolishing RASSF6 effects on Bcl-xL and apoptosis.\",\n      \"evidence\": \"OE/siRNA, YAP rescue, JC-1, Annexin V/PI in bladder cancer cells\",\n      \"pmids\": [\"31807003\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Context dependence of RASSF6-YAP regulation across tissues unmapped\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Extended miRNA-mediated silencing to miR-496, which directly represses RASSF6 to activate Wnt signaling in colorectal cancer.\",\n      \"evidence\": \"Luciferase reporter, miR-496 mimic/inhibitor, RASSF6 siRNA rescue, migration/EMT assays\",\n      \"pmids\": [\"31273789\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Link between RASSF6 loss and Wnt activation not mechanistically dissected\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Reported a context where RASSF6 promotes tumor progression by driving ubiquitination-dependent degradation of TRIM16 in esophageal carcinoma.\",\n      \"evidence\": \"IHC, OE/KD, ubiquitination assay, xenograft and metastasis models\",\n      \"pmids\": [\"31812473\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Apparent oncogenic role conflicts with tumor-suppressor findings elsewhere\", \"Mechanism of TRIM16 ubiquitin-targeting unclear\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Showed UNC119A bridges activated KRAS to RASSF6 and enhances the RASSF6-MDM2 interaction to drive p53-dependent apoptosis, connecting RAS sensing to p53 stabilization.\",\n      \"evidence\": \"Co-IP, immunofluorescence, RNAi, soft-agar, migration/invasion assays in KRAS-mutant cells\",\n      \"pmids\": [\"32554467\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Structural basis of the KRAS-UNC119A-RASSF6 assembly unknown\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Dissected the DNA-damage-triggered nuclear program, showing CDK9-phosphorylated BAF53 drives RASSF6 nuclear accumulation and that RASSF6 bridges BAF53-BAF60a-p53 to promote p53 target transcription.\",\n      \"evidence\": \"Fractionation, NLS/NES mapping, Co-IP, CDK9 inhibitor, BAF53/BAF60a siRNA, p53 target and apoptosis assays\",\n      \"pmids\": [\"34898277\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct chromatin recruitment to p53 target promoters not shown\", \"Generality across damage types unclear\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Provided in vivo genetic confirmation that Rassf6 is required for UV-induced p53 responses, DNA repair, and senescence and restrains KRAS-driven transformation and skin inflammation.\",\n      \"evidence\": \"CRISPR/Cas9 knockout mice and MEFs, RNA-seq, soft-agar, DMBA-induced inflammation\",\n      \"pmids\": [\"34652874\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Tissue-specific contributions not separated\", \"Enhanced NF-kB signaling mechanism not defined\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How RASSF6 reconciles its dual reported roles—tumor suppressor (MST2/p53/pRb/JNK) versus context-specific oncogenic TRIM16 degradation—and what determines its choice between Hippo, p53, and JNK outputs remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural model of RASSF6 multi-partner assemblies\", \"Determinants of context-dependent output unknown\", \"Direct biochemical activity (E3 cofactor vs scaffold) undefined\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [3, 17, 16]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [1, 3, 9]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [17]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [17]},\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [2]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": []}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"MST2\", \"MDM2\", \"MAGI1\", \"BCL2L1\", \"BAF53\", \"SMARCD1\", \"KRAS\", \"UNC119\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":8,"faith_total":8,"faith_pct":100.0}}