{"gene":"RASSF7","run_date":"2026-06-10T06:43:36","timeline":{"discoveries":[{"year":2008,"finding":"Xenopus RASSF7 localizes to the centrosome in a microtubule-dependent manner and is required for mitotic spindle formation; knockdown causes failure to form a mitotic spindle, mitotic arrest, nuclear breakdown, and apoptosis in the neural tube.","method":"Morpholino knockdown in Xenopus embryos, immunofluorescence localization, microtubule-dependency assay","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — loss-of-function with defined cellular phenotype (spindle failure, apoptosis), direct localization experiment with functional consequence, replicated in human cells by subsequent study","pmids":["18272789"],"is_preprint":false},{"year":2010,"finding":"Human RASSF7 localizes to the centrosome, regulates microtubule dynamics (shown by microtubule-regrowth assays), and is required for Aurora B kinase activation, chromosomal congression, and spindle formation during mitosis; knockdown inhibits cell growth and induces mitotic defects.","method":"siRNA knockdown in human cell lines, immunofluorescence, microtubule-regrowth assays, mitotic signaling kinase activity assays","journal":"The Biochemical journal","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (KD phenotype, localization, kinase activity assay, microtubule regrowth), independently replicates Xenopus findings in human cells","pmids":["20629633"],"is_preprint":false},{"year":2010,"finding":"RASSF7 interacts with GTP-bound N-Ras via its RA domain and with MKK7 to negatively regulate JNK signaling: RASSF7 promotes the phosphorylated state of MKK7 while inhibiting its ability to activate JNK, thereby suppressing stress-induced apoptosis. Under prolonged stress, RASSF7 is degraded via the ubiquitin-proteasome pathway, releasing the JNK pathway to proceed.","method":"Co-immunoprecipitation, RNAi knockdown, kinase activity assays, domain mutant analysis (RA domain), proteasome inhibitor experiments","journal":"Cell death and differentiation","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP identifying binding partners, domain mutagenesis (RA domain), kinase assay, and proteasomal degradation demonstrated with multiple orthogonal methods in a single study","pmids":["21278800"],"is_preprint":false},{"year":2015,"finding":"The coiled-coil domain of RASSF7 is necessary and sufficient for centrosomal localization, while the RA domain does not mediate localization. Truncation of the C-terminus causes RASSF7 to accumulate at the centrosome, drive centrosome defects (accumulation of γ-tubulin and amplification of γ-tubulin foci), and ultimately induce cell death; these effects require the coiled-coil-mediated centrosomal localization.","method":"Truncation construct expression in Xenopus embryos, quantitative immunofluorescence of centrosomal markers, domain deletion analysis","journal":"Developmental biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — systematic domain truncation analysis with quantitative phenotypic readout in a single lab, Xenopus system","pmids":["26569555"],"is_preprint":false},{"year":2016,"finding":"DISC1 directly associates with RASSF7 to activate the RAS/MEK/ERK signaling pathway, promoting astrogenesis; the pERK complex undergoes nuclear translocation and influences expression of astrogenesis-related genes.","method":"Co-immunoprecipitation, in vivo and in vitro knockdown/overexpression of DISC1, rescue experiments in mouse embryonic brain","journal":"Development (Cambridge, England)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP demonstrating direct association, functional rescue experiments, signaling pathway placement, single lab","pmids":["27287808"],"is_preprint":false},{"year":2018,"finding":"RASSF7 interacts with c-Myc via its RA and leucine zipper (LZ) domains, destabilizes c-Myc by promoting Cullin4B-mediated polyubiquitination and proteasomal degradation, competes with MAX for c-Myc heterodimerization, and attenuates c-Myc occupancy on target gene promoters, thereby inhibiting oncogenic transformation.","method":"Co-immunoprecipitation, domain mapping (RA and LZ constructs), ubiquitination assay, chromatin immunoprecipitation, cell transformation assay in HEK293T and HeLa cells","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — multiple orthogonal methods (Co-IP, ubiquitination assay, ChIP, domain mutants, transformation assay) in a single rigorous study establishing trimodal regulation","pmids":["30139745"],"is_preprint":false},{"year":2018,"finding":"RASSF7 promotes HCC cell proliferation by activating the MEK1/2-ERK1/2 signaling pathway; overexpression drives G1-S cell cycle transition and inhibits apoptosis, while knockdown causes G1-S arrest and apoptosis.","method":"Overexpression and siRNA knockdown in HCC cell lines, cell cycle analysis, apoptosis assay, Western blot of MEK/ERK phosphorylation","journal":"Cellular and molecular biology (Noisy-le-Grand, France)","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — KD/OE with defined phenotype and pathway placement via phospho-Western, single lab, no direct binding assay for MEK1/2","pmids":["29729697"],"is_preprint":false},{"year":1994,"finding":"HRC1 (RASSF7) maps to human chromosome 11p15.5, approximately 30 kb upstream of HRAS1, divergently transcribed from that locus; physical mapping confirmed the gene order HRC1-HRAS1-RNH within an 80-kb genomic region.","method":"Pulsed-field gel electrophoresis, Southern blot, P1 bacteriophage clone physical mapping","journal":"Genetic analysis, techniques and applications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct physical mapping with multiple complementary genomic methods, establishing chromosomal locus","pmids":["7710782"],"is_preprint":false}],"current_model":"RASSF7 is a centrosome-associated protein (localization mediated by its coiled-coil domain) that functions as a RAS effector: it binds GTP-bound N-Ras via its RA domain to suppress pro-apoptotic MKK7/JNK signaling during acute stress (while itself being degraded by the ubiquitin-proteasome pathway under prolonged stress), activates MEK/ERK signaling (including in a complex with DISC1 to promote astrogenesis), inhibits c-Myc by promoting Cullin4B-mediated ubiquitination/degradation and competing with MAX for c-Myc heterodimerization, and is required for microtubule dynamics, Aurora B activation, chromosomal congression, and mitotic spindle formation."},"narrative":{"mechanistic_narrative":"RASSF7 is a centrosome-associated RAS-family effector that couples microtubule organization to mitotic progression and stress signaling [PMID:18272789, PMID:20629633]. Localization to the centrosome is microtubule-dependent and is conferred specifically by its coiled-coil domain, which is necessary and sufficient for targeting; disrupting this organization through C-terminal truncation drives γ-tubulin accumulation, centrosome amplification, and cell death [PMID:26569555]. At the centrosome RASSF7 regulates microtubule dynamics and is required for Aurora B activation, chromosomal congression, and mitotic spindle formation, with loss of function causing spindle failure, mitotic arrest, and apoptosis [PMID:18272789, PMID:20629633]. As a RAS effector, RASSF7 binds GTP-loaded N-Ras through its RA domain and engages MKK7 to suppress pro-apoptotic JNK signaling during acute stress, while prolonged stress triggers its ubiquitin-proteasome-mediated degradation to release the pathway [PMID:21278800]. It also positively feeds MEK/ERK signaling—acting in association with DISC1 to promote astrogenesis through nuclear translocation of pERK—and activates MEK1/2-ERK1/2 to drive G1-S transition [PMID:27287808, PMID:29729697]. Through its RA and leucine-zipper domains RASSF7 antagonizes c-Myc, promoting Cullin4B-mediated polyubiquitination and degradation, competing with MAX for heterodimerization, and reducing c-Myc promoter occupancy to inhibit oncogenic transformation [PMID:30139745].","teleology":[{"year":1994,"claim":"Before any function was known, the genomic context of RASSF7 was defined, placing it immediately upstream of and divergently transcribed from HRAS1, anchoring its later identification as a RAS-pathway gene.","evidence":"Pulsed-field gel electrophoresis, Southern blot, and P1 clone physical mapping of human chromosome 11p15.5","pmids":["7710782"],"confidence":"Medium","gaps":["Establishes only chromosomal locus and gene order, no functional or biochemical role","Physical proximity to HRAS1 does not establish a regulatory or effector relationship"]},{"year":2008,"claim":"The first functional study established RASSF7 as a centrosomal protein essential for mitotic spindle formation, answering whether it has a role in cell division.","evidence":"Morpholino knockdown and immunofluorescence in Xenopus embryos with microtubule-dependency assay","pmids":["18272789"],"confidence":"High","gaps":["Mechanism by which RASSF7 promotes spindle assembly not defined","Molecular partners at the centrosome not identified"]},{"year":2010,"claim":"The mitotic role was extended to human cells and linked to specific machinery, showing RASSF7 regulates microtubule dynamics and is required for Aurora B activation and chromosomal congression.","evidence":"siRNA knockdown, immunofluorescence, microtubule-regrowth assays, and kinase activity assays in human cell lines","pmids":["20629633"],"confidence":"High","gaps":["Direct link between RASSF7 and Aurora B activation mechanism unresolved","Whether RASSF7 acts as a structural scaffold or signaling intermediate at the spindle unclear"]},{"year":2010,"claim":"RASSF7 was defined as a bona fide RAS effector that gates stress-induced apoptosis, answering how it connects RAS to cell death decisions.","evidence":"Reciprocal Co-IP, RA-domain mutagenesis, kinase assays, and proteasome inhibitor experiments","pmids":["21278800"],"confidence":"High","gaps":["Ubiquitin ligase responsible for stress-induced RASSF7 degradation not identified","Mechanism by which RASSF7 uncouples MKK7 phosphorylation from JNK activation not structurally defined"]},{"year":2015,"claim":"Domain dissection resolved which region targets RASSF7 to the centrosome, showing the coiled-coil domain is necessary and sufficient and that mislocalization causes centrosome defects.","evidence":"Truncation construct expression and quantitative centrosomal-marker immunofluorescence in Xenopus embryos","pmids":["26569555"],"confidence":"Medium","gaps":["Centrosomal docking partner of the coiled-coil domain not identified","Single-lab Xenopus system; human cell confirmation of truncation phenotype absent"]},{"year":2016,"claim":"A positive signaling arm was added, showing RASSF7 partners with DISC1 to activate RAS/MEK/ERK and drive astrogenesis, distinct from its apoptosis-suppressing role.","evidence":"Co-IP, in vivo/in vitro DISC1 perturbation, and rescue experiments in mouse embryonic brain","pmids":["27287808"],"confidence":"Medium","gaps":["Whether RASSF7 directly engages MEK/ERK components or acts via N-Ras not resolved","Single-lab finding; reciprocal validation of the DISC1-RASSF7 association limited"]},{"year":2018,"claim":"RASSF7 was shown to negatively regulate c-Myc through a trimodal mechanism, establishing a tumor-suppressive function opposed to its proliferative MEK/ERK role.","evidence":"Co-IP, RA/LZ domain mapping, ubiquitination assay, ChIP, and transformation assay in HEK293T and HeLa cells","pmids":["30139745"],"confidence":"High","gaps":["How RASSF7 recruits Cullin4B to c-Myc not defined","Reconciliation of c-Myc-inhibitory versus proliferation-promoting activities in the same cell context not addressed"]},{"year":2018,"claim":"A pro-proliferative function in hepatocellular carcinoma was demonstrated, showing RASSF7 drives G1-S transition via MEK1/2-ERK1/2 activation.","evidence":"Overexpression/knockdown in HCC cell lines with cell cycle, apoptosis, and phospho-MEK/ERK Western analyses","pmids":["29729697"],"confidence":"Medium","gaps":["No direct binding assay linking RASSF7 to MEK1/2","Single-lab study; context-dependence relative to its c-Myc-inhibitory tumor-suppressor role unresolved"]},{"year":null,"claim":"How RASSF7's centrosomal/mitotic function, RAS-effector stress signaling, MEK/ERK-promoting activity, and c-Myc-inhibitory tumor-suppression are integrated within a single cell type remains unresolved.","evidence":"","pmids":[],"confidence":"Low","gaps":["No unifying model reconciling pro-proliferative and tumor-suppressive activities","Centrosomal docking partner and structural basis of coiled-coil targeting unknown","Endogenous regulation switching RASSF7 between signaling modes not defined"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[2,5]},{"term_id":"GO:0060089","term_label":"molecular transducer activity","supporting_discovery_ids":[2,4]},{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[0,1]}],"localization":[{"term_id":"GO:0005815","term_label":"microtubule organizing center","supporting_discovery_ids":[0,1,3]},{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[0,1]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[2,4,6]},{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[0,1]},{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[2]},{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[2,5]}],"complexes":[],"partners":["NRAS","MKK7","DISC1","MYC","CUL4B","MAX"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q02833","full_name":"Ras association domain-containing protein 7","aliases":["HRAS1-related cluster protein 1"],"length_aa":373,"mass_kda":39.9,"function":"Negatively regulates stress-induced JNK activation and apoptosis by promoting MAP2K7 phosphorylation and inhibiting its ability to activate JNK. Following prolonged stress, anti-apoptotic effect stops because of degradation of RASSF7 protein via the ubiquitin-proteasome pathway. Required for the activation of AURKB and chromosomal congression during mitosis where it stimulates microtubule polymerization","subcellular_location":"Cytoplasm, cytoskeleton, microtubule organizing center, centrosome","url":"https://www.uniprot.org/uniprotkb/Q02833/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/RASSF7","classification":"Not Classified","n_dependent_lines":2,"n_total_lines":1208,"dependency_fraction":0.0016556291390728477},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/RASSF7","total_profiled":1310},"omim":[{"mim_id":"614713","title":"RAS ASSOCIATION DOMAIN FAMILY, MEMBER 10; RASSF10","url":"https://www.omim.org/entry/614713"},{"mim_id":"610383","title":"RAS ASSOCIATION DOMAIN FAMILY, MEMBER 9; RASSF9","url":"https://www.omim.org/entry/610383"},{"mim_id":"143023","title":"RAS ASSOCIATION DOMAIN FAMILY, MEMBER 7; RASSF7","url":"https://www.omim.org/entry/143023"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Centriolar satellite","reliability":"Supported"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"kidney","ntpm":121.5}],"url":"https://www.proteinatlas.org/search/RASSF7"},"hgnc":{"alias_symbol":["HRC1","HRAS1","FAP88","CFAP88"],"prev_symbol":["C11orf13"]},"alphafold":{"accession":"Q02833","domains":[{"cath_id":"3.10.20.90","chopping":"7-87","consensus_level":"high","plddt":91.1148,"start":7,"end":87},{"cath_id":"1.20.5","chopping":"152-227","consensus_level":"medium","plddt":93.2595,"start":152,"end":227},{"cath_id":"1.20.5","chopping":"230-280","consensus_level":"medium","plddt":92.2773,"start":230,"end":280}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q02833","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q02833-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q02833-F1-predicted_aligned_error_v6.png","plddt_mean":73.75},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=RASSF7","jax_strain_url":"https://www.jax.org/strain/search?query=RASSF7"},"sequence":{"accession":"Q02833","fasta_url":"https://rest.uniprot.org/uniprotkb/Q02833.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q02833/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q02833"}},"corpus_meta":[{"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":"21116130","id":"PMC_21116130","title":"N-terminal RASSF family: RASSF7-RASSF10.","date":"2011","source":"Epigenetics","url":"https://pubmed.ncbi.nlm.nih.gov/21116130","citation_count":60,"is_preprint":false},{"pmid":"18272789","id":"PMC_18272789","title":"RASSF7 is a member of a new family of RAS association domain-containing proteins and is required for completing mitosis.","date":"2008","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/18272789","citation_count":50,"is_preprint":false},{"pmid":"21278800","id":"PMC_21278800","title":"RASSF7 negatively regulates pro-apoptotic JNK signaling by inhibiting the activity of phosphorylated-MKK7.","date":"2010","source":"Cell death and differentiation","url":"https://pubmed.ncbi.nlm.nih.gov/21278800","citation_count":34,"is_preprint":false},{"pmid":"20629633","id":"PMC_20629633","title":"Human RASSF7 regulates the microtubule cytoskeleton and is required for spindle formation, Aurora B activation and chromosomal congression during mitosis.","date":"2010","source":"The Biochemical journal","url":"https://pubmed.ncbi.nlm.nih.gov/20629633","citation_count":24,"is_preprint":false},{"pmid":"27287808","id":"PMC_27287808","title":"DISC1 regulates astrogenesis in the embryonic brain via modulation of RAS/MEK/ERK signaling through RASSF7.","date":"2016","source":"Development (Cambridge, England)","url":"https://pubmed.ncbi.nlm.nih.gov/27287808","citation_count":23,"is_preprint":false},{"pmid":"26569555","id":"PMC_26569555","title":"Truncated RASSF7 promotes centrosomal defects and cell death.","date":"2015","source":"Developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/26569555","citation_count":7,"is_preprint":false},{"pmid":"26884887","id":"PMC_26884887","title":"RASSF7 expression and its regulatory roles on apoptosis in human intervertebral disc degeneration.","date":"2015","source":"International journal of clinical and experimental pathology","url":"https://pubmed.ncbi.nlm.nih.gov/26884887","citation_count":7,"is_preprint":false},{"pmid":"30139745","id":"PMC_30139745","title":"The non-enzymatic RAS effector RASSF7 inhibits oncogenic c-Myc function.","date":"2018","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/30139745","citation_count":7,"is_preprint":false},{"pmid":"34217978","id":"PMC_34217978","title":"Sonapatha (Oroxylum indicum) mediates cytotoxicity in cultured HeLa cells by inducing apoptosis and suppressing NF-κB, COX-2, RASSF7 and NRF2.","date":"2021","source":"Bioorganic chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/34217978","citation_count":7,"is_preprint":false},{"pmid":"29729697","id":"PMC_29729697","title":"RASSF7 promotes cell proliferation through activating MEK1/2-ERK1/2 signaling pathway in hepatocellular carcinoma.","date":"2018","source":"Cellular and molecular biology (Noisy-le-Grand, France)","url":"https://pubmed.ncbi.nlm.nih.gov/29729697","citation_count":5,"is_preprint":false},{"pmid":"24516056","id":"PMC_24516056","title":"RASSF7 and RASSF8 proteins are predictive factors for development and metastasis in malignant thyroid neoplasms.","date":"2013","source":"Journal of cancer research and therapeutics","url":"https://pubmed.ncbi.nlm.nih.gov/24516056","citation_count":4,"is_preprint":false},{"pmid":"36200241","id":"PMC_36200241","title":"A Number of the N-terminal RASSF Family: RASSF7.","date":"2024","source":"Anti-cancer agents in medicinal chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/36200241","citation_count":4,"is_preprint":false},{"pmid":"7710782","id":"PMC_7710782","title":"A single P1 clone bearing three genes from human chromosome 11p15.5: HRC1, HRAS1, and RNH.","date":"1994","source":"Genetic analysis, techniques and applications","url":"https://pubmed.ncbi.nlm.nih.gov/7710782","citation_count":3,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":8665,"output_tokens":2108,"usd":0.028807,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":9091,"output_tokens":3095,"usd":0.061415,"stage2_stop_reason":"end_turn"},"total_usd":0.090222,"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\": 2008,\n      \"finding\": \"Xenopus RASSF7 localizes to the centrosome in a microtubule-dependent manner and is required for mitotic spindle formation; knockdown causes failure to form a mitotic spindle, mitotic arrest, nuclear breakdown, and apoptosis in the neural tube.\",\n      \"method\": \"Morpholino knockdown in Xenopus embryos, immunofluorescence localization, microtubule-dependency assay\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — loss-of-function with defined cellular phenotype (spindle failure, apoptosis), direct localization experiment with functional consequence, replicated in human cells by subsequent study\",\n      \"pmids\": [\"18272789\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Human RASSF7 localizes to the centrosome, regulates microtubule dynamics (shown by microtubule-regrowth assays), and is required for Aurora B kinase activation, chromosomal congression, and spindle formation during mitosis; knockdown inhibits cell growth and induces mitotic defects.\",\n      \"method\": \"siRNA knockdown in human cell lines, immunofluorescence, microtubule-regrowth assays, mitotic signaling kinase activity assays\",\n      \"journal\": \"The Biochemical journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (KD phenotype, localization, kinase activity assay, microtubule regrowth), independently replicates Xenopus findings in human cells\",\n      \"pmids\": [\"20629633\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"RASSF7 interacts with GTP-bound N-Ras via its RA domain and with MKK7 to negatively regulate JNK signaling: RASSF7 promotes the phosphorylated state of MKK7 while inhibiting its ability to activate JNK, thereby suppressing stress-induced apoptosis. Under prolonged stress, RASSF7 is degraded via the ubiquitin-proteasome pathway, releasing the JNK pathway to proceed.\",\n      \"method\": \"Co-immunoprecipitation, RNAi knockdown, kinase activity assays, domain mutant analysis (RA domain), proteasome inhibitor experiments\",\n      \"journal\": \"Cell death and differentiation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP identifying binding partners, domain mutagenesis (RA domain), kinase assay, and proteasomal degradation demonstrated with multiple orthogonal methods in a single study\",\n      \"pmids\": [\"21278800\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"The coiled-coil domain of RASSF7 is necessary and sufficient for centrosomal localization, while the RA domain does not mediate localization. Truncation of the C-terminus causes RASSF7 to accumulate at the centrosome, drive centrosome defects (accumulation of γ-tubulin and amplification of γ-tubulin foci), and ultimately induce cell death; these effects require the coiled-coil-mediated centrosomal localization.\",\n      \"method\": \"Truncation construct expression in Xenopus embryos, quantitative immunofluorescence of centrosomal markers, domain deletion analysis\",\n      \"journal\": \"Developmental biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — systematic domain truncation analysis with quantitative phenotypic readout in a single lab, Xenopus system\",\n      \"pmids\": [\"26569555\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"DISC1 directly associates with RASSF7 to activate the RAS/MEK/ERK signaling pathway, promoting astrogenesis; the pERK complex undergoes nuclear translocation and influences expression of astrogenesis-related genes.\",\n      \"method\": \"Co-immunoprecipitation, in vivo and in vitro knockdown/overexpression of DISC1, rescue experiments in mouse embryonic brain\",\n      \"journal\": \"Development (Cambridge, England)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP demonstrating direct association, functional rescue experiments, signaling pathway placement, single lab\",\n      \"pmids\": [\"27287808\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"RASSF7 interacts with c-Myc via its RA and leucine zipper (LZ) domains, destabilizes c-Myc by promoting Cullin4B-mediated polyubiquitination and proteasomal degradation, competes with MAX for c-Myc heterodimerization, and attenuates c-Myc occupancy on target gene promoters, thereby inhibiting oncogenic transformation.\",\n      \"method\": \"Co-immunoprecipitation, domain mapping (RA and LZ constructs), ubiquitination assay, chromatin immunoprecipitation, cell transformation assay in HEK293T and HeLa cells\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — multiple orthogonal methods (Co-IP, ubiquitination assay, ChIP, domain mutants, transformation assay) in a single rigorous study establishing trimodal regulation\",\n      \"pmids\": [\"30139745\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"RASSF7 promotes HCC cell proliferation by activating the MEK1/2-ERK1/2 signaling pathway; overexpression drives G1-S cell cycle transition and inhibits apoptosis, while knockdown causes G1-S arrest and apoptosis.\",\n      \"method\": \"Overexpression and siRNA knockdown in HCC cell lines, cell cycle analysis, apoptosis assay, Western blot of MEK/ERK phosphorylation\",\n      \"journal\": \"Cellular and molecular biology (Noisy-le-Grand, France)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — KD/OE with defined phenotype and pathway placement via phospho-Western, single lab, no direct binding assay for MEK1/2\",\n      \"pmids\": [\"29729697\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1994,\n      \"finding\": \"HRC1 (RASSF7) maps to human chromosome 11p15.5, approximately 30 kb upstream of HRAS1, divergently transcribed from that locus; physical mapping confirmed the gene order HRC1-HRAS1-RNH within an 80-kb genomic region.\",\n      \"method\": \"Pulsed-field gel electrophoresis, Southern blot, P1 bacteriophage clone physical mapping\",\n      \"journal\": \"Genetic analysis, techniques and applications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct physical mapping with multiple complementary genomic methods, establishing chromosomal locus\",\n      \"pmids\": [\"7710782\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"RASSF7 is a centrosome-associated protein (localization mediated by its coiled-coil domain) that functions as a RAS effector: it binds GTP-bound N-Ras via its RA domain to suppress pro-apoptotic MKK7/JNK signaling during acute stress (while itself being degraded by the ubiquitin-proteasome pathway under prolonged stress), activates MEK/ERK signaling (including in a complex with DISC1 to promote astrogenesis), inhibits c-Myc by promoting Cullin4B-mediated ubiquitination/degradation and competing with MAX for c-Myc heterodimerization, and is required for microtubule dynamics, Aurora B activation, chromosomal congression, and mitotic spindle formation.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"RASSF7 is a centrosome-associated RAS-family effector that couples microtubule organization to mitotic progression and stress signaling [#0, #1]. Localization to the centrosome is microtubule-dependent and is conferred specifically by its coiled-coil domain, which is necessary and sufficient for targeting; disrupting this organization through C-terminal truncation drives γ-tubulin accumulation, centrosome amplification, and cell death [#3]. At the centrosome RASSF7 regulates microtubule dynamics and is required for Aurora B activation, chromosomal congression, and mitotic spindle formation, with loss of function causing spindle failure, mitotic arrest, and apoptosis [#0, #1]. As a RAS effector, RASSF7 binds GTP-loaded N-Ras through its RA domain and engages MKK7 to suppress pro-apoptotic JNK signaling during acute stress, while prolonged stress triggers its ubiquitin-proteasome-mediated degradation to release the pathway [#2]. It also positively feeds MEK/ERK signaling—acting in association with DISC1 to promote astrogenesis through nuclear translocation of pERK—and activates MEK1/2-ERK1/2 to drive G1-S transition [#4, #6]. Through its RA and leucine-zipper domains RASSF7 antagonizes c-Myc, promoting Cullin4B-mediated polyubiquitination and degradation, competing with MAX for heterodimerization, and reducing c-Myc promoter occupancy to inhibit oncogenic transformation [#5].\",\n  \"teleology\": [\n    {\n      \"year\": 1994,\n      \"claim\": \"Before any function was known, the genomic context of RASSF7 was defined, placing it immediately upstream of and divergently transcribed from HRAS1, anchoring its later identification as a RAS-pathway gene.\",\n      \"evidence\": \"Pulsed-field gel electrophoresis, Southern blot, and P1 clone physical mapping of human chromosome 11p15.5\",\n      \"pmids\": [\"7710782\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Establishes only chromosomal locus and gene order, no functional or biochemical role\", \"Physical proximity to HRAS1 does not establish a regulatory or effector relationship\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"The first functional study established RASSF7 as a centrosomal protein essential for mitotic spindle formation, answering whether it has a role in cell division.\",\n      \"evidence\": \"Morpholino knockdown and immunofluorescence in Xenopus embryos with microtubule-dependency assay\",\n      \"pmids\": [\"18272789\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism by which RASSF7 promotes spindle assembly not defined\", \"Molecular partners at the centrosome not identified\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"The mitotic role was extended to human cells and linked to specific machinery, showing RASSF7 regulates microtubule dynamics and is required for Aurora B activation and chromosomal congression.\",\n      \"evidence\": \"siRNA knockdown, immunofluorescence, microtubule-regrowth assays, and kinase activity assays in human cell lines\",\n      \"pmids\": [\"20629633\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct link between RASSF7 and Aurora B activation mechanism unresolved\", \"Whether RASSF7 acts as a structural scaffold or signaling intermediate at the spindle unclear\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"RASSF7 was defined as a bona fide RAS effector that gates stress-induced apoptosis, answering how it connects RAS to cell death decisions.\",\n      \"evidence\": \"Reciprocal Co-IP, RA-domain mutagenesis, kinase assays, and proteasome inhibitor experiments\",\n      \"pmids\": [\"21278800\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Ubiquitin ligase responsible for stress-induced RASSF7 degradation not identified\", \"Mechanism by which RASSF7 uncouples MKK7 phosphorylation from JNK activation not structurally defined\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Domain dissection resolved which region targets RASSF7 to the centrosome, showing the coiled-coil domain is necessary and sufficient and that mislocalization causes centrosome defects.\",\n      \"evidence\": \"Truncation construct expression and quantitative centrosomal-marker immunofluorescence in Xenopus embryos\",\n      \"pmids\": [\"26569555\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Centrosomal docking partner of the coiled-coil domain not identified\", \"Single-lab Xenopus system; human cell confirmation of truncation phenotype absent\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"A positive signaling arm was added, showing RASSF7 partners with DISC1 to activate RAS/MEK/ERK and drive astrogenesis, distinct from its apoptosis-suppressing role.\",\n      \"evidence\": \"Co-IP, in vivo/in vitro DISC1 perturbation, and rescue experiments in mouse embryonic brain\",\n      \"pmids\": [\"27287808\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether RASSF7 directly engages MEK/ERK components or acts via N-Ras not resolved\", \"Single-lab finding; reciprocal validation of the DISC1-RASSF7 association limited\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"RASSF7 was shown to negatively regulate c-Myc through a trimodal mechanism, establishing a tumor-suppressive function opposed to its proliferative MEK/ERK role.\",\n      \"evidence\": \"Co-IP, RA/LZ domain mapping, ubiquitination assay, ChIP, and transformation assay in HEK293T and HeLa cells\",\n      \"pmids\": [\"30139745\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How RASSF7 recruits Cullin4B to c-Myc not defined\", \"Reconciliation of c-Myc-inhibitory versus proliferation-promoting activities in the same cell context not addressed\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"A pro-proliferative function in hepatocellular carcinoma was demonstrated, showing RASSF7 drives G1-S transition via MEK1/2-ERK1/2 activation.\",\n      \"evidence\": \"Overexpression/knockdown in HCC cell lines with cell cycle, apoptosis, and phospho-MEK/ERK Western analyses\",\n      \"pmids\": [\"29729697\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No direct binding assay linking RASSF7 to MEK1/2\", \"Single-lab study; context-dependence relative to its c-Myc-inhibitory tumor-suppressor role unresolved\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How RASSF7's centrosomal/mitotic function, RAS-effector stress signaling, MEK/ERK-promoting activity, and c-Myc-inhibitory tumor-suppression are integrated within a single cell type remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No unifying model reconciling pro-proliferative and tumor-suppressive activities\", \"Centrosomal docking partner and structural basis of coiled-coil targeting unknown\", \"Endogenous regulation switching RASSF7 between signaling modes not defined\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [2, 5]},\n      {\"term_id\": \"GO:0060089\", \"supporting_discovery_ids\": [2, 4]},\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [0, 1]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005815\", \"supporting_discovery_ids\": [0, 1, 3]},\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [0, 1]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [2, 4, 6]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [0, 1]},\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [2]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [2, 5]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"NRAS\", \"MKK7\", \"DISC1\", \"MYC\", \"CUL4B\", \"MAX\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}