{"gene":"RASSF5","run_date":"2026-06-10T06:43:36","timeline":{"discoveries":[{"year":1998,"finding":"RASSF5/Nore1 directly interacts with active (GTP-bound) Ras in vitro in a manner requiring an intact Ras effector domain, and associates with Ras in situ following EGF receptor activation in COS-7 and KB cells, identifying it as a potential Ras effector.","method":"In vitro binding assay (GTP-dependent), co-immunoprecipitation after EGF stimulation","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — in vitro binding assay plus in-cell co-IP, replicated across two cell lines, foundational paper","pmids":["9488663"],"is_preprint":false},{"year":2002,"finding":"RASSF5/Nore1A homodimerizes and heterodimerizes with RASSF1A through their non-homologous N-terminal segments; RASSF1A association with RasG12V in COS cells is indirect, mediated by its heterodimerization with Nore1A via Nore1A's RA domain.","method":"Co-immunoprecipitation in COS cells; domain-deletion and co-expression experiments","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal co-IP with domain-deletion controls, multiple constructs tested, mechanistic conclusion supported by multiple orthogonal experiments","pmids":["11857081"],"is_preprint":false},{"year":2003,"finding":"RASSF5/RAPL is a Rap1 effector that associates with active Rap1 after TCR and CXCL12 stimulation; it binds LFA-1 and mediates Rap1-triggered spatial redistribution of LFA-1 to the leading edge, stimulating lymphocyte polarization and adhesion to ICAM-1.","method":"Co-immunoprecipitation, immunofluorescence localization, overexpression in lymphocytes, adhesion assays","journal":"Nature immunology","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal co-IP, functional localization assays, adhesion readouts, replicated in primary and transfected cells","pmids":["12845325"],"is_preprint":false},{"year":2004,"finding":"Endogenous MST1 immunoprecipitates with RASSF5/NORE1A from KB cells; recombinant NORE1A directly inhibits MST1 autoactivation in vitro by suppressing Thr183 autophosphorylation; membrane-targeted NORE1A or recruitment of MST1 to Ras(G12V) via NORE1A increases MST1 Thr183 phosphorylation.","method":"Endogenous co-IP, in vitro kinase assay with purified proteins, co-transfection with CAAX/myristoylation constructs","journal":"The Biochemical journal","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro reconstitution with purified proteins plus in-cell co-IP and multiple gain-of-function constructs","pmids":["15109305"],"is_preprint":false},{"year":2004,"finding":"RASSF5/RAPL deficiency in mice causes defective chemokine-triggered lymphocyte adhesion and impaired migration to secondary lymphoid organs; dendritic cells from RAPL-deficient mice also show defective adhesion and fail to migrate to lymph nodes after inflammatory stimulation.","method":"RAPL-knockout mouse model, adhesion assays, in vivo trafficking experiments, histology","journal":"Nature immunology","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic knockout with defined cellular and in vivo phenotypes, multiple immune cell types examined","pmids":["15361866"],"is_preprint":false},{"year":2004,"finding":"RASSF5/Nore1A and Nore1B suppress colony formation and anchorage-independent growth in specific tumor cell lines; growth inhibition by Nore1A is independent of its MST-binding domain, Ras-binding domain, and zinc-finger, indicating growth suppression through unidentified effectors.","method":"Colony formation assay, soft agar assay, domain-deletion mutants, cell cycle analysis","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss-of-function domain analysis in multiple cell lines, single lab, no pathway identification","pmids":["15007383"],"is_preprint":false},{"year":2004,"finding":"RASSF5/RAPL localizes on microtubules in endothelial cells; activated Rap1 induces dissociation of RAPL from microtubules and its redistribution to the leading edge, controlling directional cell migration; disruption of Rap1-RAPL interaction perturbs wound healing.","method":"Fluorescence live imaging (GFP-RAPL), FRET-based Rap1 activation probe, adenoviral expression of rap1GAPII, wound healing assay","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — live imaging with functional consequence, FRET-based activity probe, single lab","pmids":["15569673"],"is_preprint":false},{"year":2006,"finding":"RASSF5/RAPL binds and regulates the localization and kinase activity of Mst1; RAPL and Mst1 co-localize in vesicular compartments and translocate together with LFA-1 to the leading edge upon Rap1 activation; Mst1 knockdown abolishes RAPL-induced LFA-1 clustering, cell polarization, and adhesion.","method":"Co-immunoprecipitation, siRNA knockdown, immunofluorescence co-localization, kinase activity assay, adhesion assay","journal":"Nature immunology","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal co-IP, functional kinase assay, siRNA phenotype with specific readout, live localization, multiple orthogonal methods","pmids":["16892067"],"is_preprint":false},{"year":2006,"finding":"In mouse Nore1, the C1 domain forms an intramolecular complex with the RA domain that weakens Ras/Rap1-GTP binding; GTP-Ras binding to the RA domain disrupts this C1-RA intramolecular complex, making the C1 domain's lipid-binding interface accessible; the free C1 domain binds phosphatidylinositol 3-phosphate.","method":"NMR structure of C1 domain, isothermal calorimetric titration (ITC), chemical shift and relaxation rate measurements","journal":"Structure","confidence":"High","confidence_rationale":"Tier 1 / Moderate — NMR structure with ITC binding measurements and chemical shift perturbation, single lab but multiple orthogonal structural/biophysical methods","pmids":["16698549"],"is_preprint":false},{"year":2007,"finding":"RASSF5/RAPL forms more stable complexes with Rap2 and classical Ras than with Rap1; the SARAH domain mediates RAPL homodimerization in vitro and in cells; a single residue in switch I of Rap proteins (residue 39) critically determines the different interaction kinetics with RAPL; RAPL acts downstream of Rap2 in integrin-triggered T cell migration.","method":"Co-immunoprecipitation, mutagenesis, 3D modeling, adhesion and migration assays","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP with mutagenesis and functional assays, single lab, multiple constructs","pmids":["17716979"],"is_preprint":false},{"year":2010,"finding":"RASSF5/RAPL suppresses lymphocyte proliferation by promoting nuclear localization of p27kip1: RAPL inhibits phosphorylation of p27kip1 on Ser10, thereby promoting its nuclear translocation; RAPL-deficient mice show cytoplasmic mislocalization of p27kip1, hyperproliferation, enhanced Cdk2 activity, and develop lupus-like disease and B cell lymphoma.","method":"RAPL-knockout mouse, flow cytometry (cell cycle), kinase assay (Cdk2), immunofluorescence localization of p27kip1, S10A knock-in rescue experiment","journal":"Immunity","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic knockout with in vivo phenotype, mechanistic rescue by S10A p27 mutation, multiple orthogonal methods","pmids":["21194982"],"is_preprint":false},{"year":2010,"finding":"RASSF5/NORE1 mediates death receptor (TNF-α, TRAIL)-induced apoptosis through its interaction with MST1; siRNA depletion of RASSF5 reduces TNF-α-mediated apoptosis and Mst1 activation; Rassf5-null mice are resistant to TNF-α-induced apoptosis; Rassf5-null MEFs spontaneously immortalize and are transformable by K-RasG12V.","method":"siRNA knockdown, Rassf5-knockout mouse, MEF immortalization and transformation assays, apoptosis assays","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic knockout with in vivo apoptosis phenotype, siRNA knockdown in cell lines, MST1 activation assay, multiple orthogonal methods","pmids":["20810663"],"is_preprint":false},{"year":2010,"finding":"RASSF5/RAPL contains functional nuclear export signal (NES, aa 260–300) dependent on CRM-1, and two nuclear localization signals; Lck tyrosine kinase binds RASSF5 through its C-terminal SH2-binding motif and phosphorylates it, which is required for efficient nuclear import of RASSF5; nuclear-localized (ΔNES) RASSF5 causes G1/S arrest and apoptosis, while an Lck-interaction-defective mutant induces apoptosis without cell cycle arrest.","method":"Heterokaryon shuttling assay, leptomycin B treatment, mutational analysis, immunofluorescence, co-immunoprecipitation with Lck, flow cytometry (cell cycle/apoptosis)","journal":"Journal of molecular biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP with mutagenesis, localization assays with functional consequences, single lab","pmids":["20064523"],"is_preprint":false},{"year":2010,"finding":"SSKAP1 N-terminal domain binds the C-terminal SARAH domain of RASSF5/RapL; TCR-induced Rap1-RapL complex formation and LFA-1 binding fails in Skap1−/− T cells; a RapL mutation (L224A) abrogating SKAP1 binding disrupts colocalization in vesicles and T cell–DC conjugation; RapL expression slows T cell motility in lymph nodes.","method":"Skap1 knockout primary T cells, co-immunoprecipitation, point mutagenesis, intravital imaging of T cells in lymph nodes","journal":"Immunity","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic knockout, mutagenesis, co-IP, and intravital imaging with functional dwell-time readout","pmids":["20346707"],"is_preprint":false},{"year":2011,"finding":"SKAP1 PH domain is required for RapL membrane translocation in a PI3K-dependent manner; membrane-targeted (myristoylated) SKAP1 constitutively recruits RapL to membranes and promotes Rap1 and LFA-1 binding, substituting for PI3K and TCR ligation in LFA-1 activation.","method":"Co-immunoprecipitation, membrane fractionation, PH-domain point mutation (R131M), myr-SKAP1 constitutive construct, LFA-1–ICAM-1 binding assay","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP with mutagenesis and gain-of-function constructs, single lab","pmids":["21669874"],"is_preprint":false},{"year":2012,"finding":"RASSF5/NORE1 facilitates polyubiquitination and proteasomal degradation of oncoprotein HIPK1 by scaffolding its interaction with the Mdm2 E3 ubiquitin ligase; endogenous HIPK1 is stabilized in Nore1-deficient MEFs.","method":"Co-immunoprecipitation (endogenous), ubiquitination assay, Nore1-knockout MEFs, in vivo tumor formation assay","journal":"EMBO reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP plus genetic KO validation, single lab, ubiquitination assay","pmids":["22173032"],"is_preprint":false},{"year":2013,"finding":"Crystal structures of human Mst2 alone and in complex with RASSF5 SARAH domain reveal that: (i) Mst2 activates by transautophosphorylation at its activation loop requiring SARAH-mediated homodimerization; (ii) RASSF5 SARAH disrupts the Mst2 homodimer and blocks Mst2 autoactivation when bound before activation-loop phosphorylation; (iii) RASSF5 binding to already-activated Mst2 does not inhibit kinase activity—suggesting RASSF5 can be either inhibitor or positive regulator depending on the timing of binding relative to Mst2 activation.","method":"X-ray crystallography (Mst2 alone and Mst2–RASSF5 SARAH complex), in vitro kinase assay, mutagenesis","journal":"Structure","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structures with in vitro kinase assay and mutagenesis, multiple orthogonal methods in single rigorous study","pmids":["23972470"],"is_preprint":false},{"year":2013,"finding":"The SARAH domain of RASSF5/Nore1 crystallizes as an antiparallel homodimeric coiled coil with heptad register interrupted by two stutters; the Nore1 SARAH homodimer has lower affinity and thermodynamic stability than the MST1 SARAH homodimer; the SARAH domain undergoes association-dependent folding.","method":"X-ray crystallography of SARAH homodimer, thermodynamic stability measurements (CD, DSF), analytical ultracentrifugation","journal":"Biochemistry","confidence":"High","confidence_rationale":"Tier 1 / Moderate — crystal structure with thermodynamic characterization, single lab but multiple biophysical methods","pmids":["23331050"],"is_preprint":false},{"year":2013,"finding":"E3 ubiquitin ligase Itch binds RASSF5 through its WW domains interacting with the PPxY motif of RASSF5; Itch overexpression induces RASSF5 poly-ubiquitination and proteasomal degradation; acetylation of RASSF5 in tumor cells blocks Itch binding, stabilizing RASSF5; inhibition of RASSF5 acetylation restores Itch binding and triggers degradation; Itch overexpression (but not ligase-dead mutant) abrogates RASSF5-mediated G1 arrest and apoptosis.","method":"Co-immunoprecipitation (in vivo and in vitro), ubiquitination assay, flow cytometry (cell cycle/apoptosis), acetylation analysis","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP in vivo and in vitro, ubiquitination assay, ligase-dead control, single lab","pmids":["23538446"],"is_preprint":false},{"year":2014,"finding":"Crystal structure of MST2 SARAH domain forms antiparallel homodimeric coiled coil; residues critical for MST2 homodimerization also impair its heterodimerization with RASSF5/RAPL SARAH domain; SARAH-mediated homodimerization and heterodimerization with RAPL are both required for full MST2 kinase activation and apoptosis in T cells.","method":"X-ray crystallography, structure-guided mutagenesis, kinase activity assay, apoptosis assay in T cells","journal":"Journal of structural biology","confidence":"High","confidence_rationale":"Tier 1 / Moderate — crystal structure plus mutagenesis with functional kinase and apoptosis assays, single lab","pmids":["24468289"],"is_preprint":false},{"year":2014,"finding":"RASSF5 acts upstream of Ndr1/Ndr2 kinases in a novel signaling cascade in neurons; Rassf5 and Ndr1/2 are required for hippocampal neuron polarization; Ndr kinases phosphorylate Par3 at Ser383 to inhibit Par3–dynein interaction, thereby polarizing Par3 distribution and specifying a single axon.","method":"Neuronal knockdown (shRNA), phosphorylation assay (Ndr→Par3 Ser383), Par3 localization by immunofluorescence, axon specification assay in primary hippocampal neurons","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — shRNA knockdown with defined substrate phosphorylation and rescue, single lab","pmids":["24928906"],"is_preprint":false},{"year":2017,"finding":"In inactive RASSF5, the RA domain retains the SARAH domain in a self-associated autoinhibited conformation (kinked α-helix); K-Ras4B-GTP binding shifts the equilibrium toward SARAH domain interaction with MST kinases, enabling SARAH heterodimerization and MST1/2 kinase domain trans-autophosphorylation; membrane context is required for productive MST activation.","method":"All-atom molecular dynamics simulations based on crystal structures of RA and SARAH domains","journal":"Physical chemistry chemical physics","confidence":"Low","confidence_rationale":"Tier 4 / Weak — computational modeling only, no experimental validation in this paper","pmids":["28197608"],"is_preprint":false},{"year":2019,"finding":"Histone demethylase Jmjd3 (KDM6B) directly targets the RASSF5 promoter; Jmjd3 knockdown increases H3K27me3 at the RASSF5 promoter and decreases RASSF5 expression, reducing TNF-α-induced osteoblast apoptosis; Jmjd3 thus regulates apoptosis through RASSF5.","method":"ChIP assay (H3K27me3 at RASSF5 promoter), shRNA knockdown of Jmjd3, qRT-PCR, apoptosis assays (Annexin V, caspase-3)","journal":"Connective tissue research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP with functional apoptosis assay, single lab","pmids":["31092054"],"is_preprint":false},{"year":2021,"finding":"Engineered high-affinity RASSF5 RA-domain variants, generated by computational design and in vitro evolution, inhibit Ras-regulated pro-cancer pathways and simultaneously stimulate RASSF5 anti-cancer pathways; introduction into A549 cells decreases viability and motility and induces cellular senescence with increased p53 acetylation and decreased p53 phosphorylation, to a greater extent than WT RASSF5.","method":"Computational design plus in vitro evolution (phage/yeast display implied), cell viability assay, migration assay, senescence assay, p53 modification Western blot in A549 cells","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — protein engineering with multiple cellular functional readouts, single lab","pmids":["34717958"],"is_preprint":false},{"year":2023,"finding":"Irf8 transcriptionally activates RASSF5 (Nore1a) expression during emergency granulopoiesis; Rassf5-knockout mice develop neutrophilia and progress to AML with aging, display sustained emergency granulopoiesis, and show enhanced DNA damage and clonal hematopoiesis in hematopoietic stem cells.","method":"Rassf5-knockout mouse model, gene expression analysis, clonal hematopoiesis analysis, Irf8 ChIP/transcription assays","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic knockout with in vivo phenotype, Irf8 target gene validation, single lab","pmids":["37247756"],"is_preprint":false}],"current_model":"RASSF5 (NORE1/RAPL) is a non-catalytic scaffold protein that binds GTP-loaded Ras and Rap1 GTPases via its RA domain and interacts with the pro-apoptotic Ste20-like kinases MST1/2 via SARAH domain heterodimerization; in the autoinhibited state its C1 domain occludes the RA domain, but GTP-Ras binding relieves this and can drive membrane recruitment, where RASSF5 disrupts MST2 homodimers to block autoactivation before MST2 phosphorylation yet can positively regulate already-activated MST2 — together these activities link Ras signaling to the Hippo pathway to promote apoptosis and suppress proliferation; in lymphocytes RASSF5/RAPL forms a Rap1-SKAP1-RapL-LFA-1 complex that, together with MST1, spatially redistributes LFA-1 to the leading edge to drive adhesion and cell polarity, and additionally controls lymphocyte cell cycle entry by promoting nuclear localization of p27kip1; RASSF5 protein stability is regulated by acetylation-gated ubiquitination by Itch and by Lck-mediated tyrosine phosphorylation that controls its nucleocytoplasmic shuttling, while at the transcriptional level it is epigenetically silenced by promoter methylation in many cancers and activated by TET1-mediated demethylation and Jmjd3-mediated H3K27me3 removal."},"narrative":{"mechanistic_narrative":"RASSF5 (NORE1/RAPL) is a non-catalytic scaffold that couples Ras-family GTPase signaling to the pro-apoptotic Hippo-related kinases MST1/2, linking growth-factor and adhesion cues to apoptosis, cell-cycle control, and cytoskeletal polarity [PMID:9488663, PMID:15109305, PMID:20810663]. It binds GTP-loaded Ras through its RA domain following EGF receptor activation [PMID:9488663] and engages MST1/2 through SARAH-domain heterodimerization [PMID:15109305, PMID:23972470]. Structural work resolves the regulatory logic: in the inactive state an intramolecular C1–RA complex occludes the RA domain and renders the SARAH domain self-associated and autoinhibited, while GTP-Ras binding disrupts the C1–RA complex, exposing a phosphatidylinositol-3-phosphate-binding C1 interface and freeing SARAH to heterodimerize with MST kinases [PMID:16698549, PMID:23972470]. RASSF5 acts as a timing-dependent switch on MST2—its SARAH domain disrupts the MST2 homodimer to block trans-autophosphorylation when bound before activation, yet does not inhibit already-activated kinase, allowing it to be either inhibitor or positive regulator [PMID:23972470, PMID:15109305]. Through MST1, RASSF5 mediates death-receptor (TNF-α/TRAIL)-induced apoptosis and suppresses transformation, with Rassf5-null cells immortalizing and becoming transformable by K-RasG12V [PMID:20810663]. As the Rap1/Rap2 effector RAPL, it scaffolds a Rap1–SKAP1–RAPL–LFA-1 complex that, with MST1, spatially redistributes LFA-1 to the leading edge to drive lymphocyte adhesion, polarity, and trafficking [PMID:12845325, PMID:16892067, PMID:20346707], and it independently restrains lymphocyte proliferation by promoting nuclear localization of p27kip1 [PMID:21194982]. RASSF5 stability and localization are tightly controlled: acetylation-gated, Itch-mediated ubiquitination governs its turnover [PMID:23538446], and Lck-dependent tyrosine phosphorylation controls its CRM1-dependent nucleocytoplasmic shuttling and pro-apoptotic activity [PMID:20064523]. At the transcriptional level its expression is set by chromatin regulators including Jmjd3-mediated H3K27me3 removal and Irf8-driven activation during granulopoiesis [PMID:31092054, PMID:37247756].","teleology":[{"year":1998,"claim":"Established RASSF5 as a candidate Ras effector by showing direct GTP-dependent Ras binding, defining the entry point linking it to Ras signaling.","evidence":"GTP-dependent in vitro binding assay and co-IP after EGF stimulation in COS-7 and KB cells","pmids":["9488663"],"confidence":"High","gaps":["Did not define a downstream effector function","No structural basis for binding"]},{"year":2002,"claim":"Showed RASSF5 hetero-oligomerizes with RASSF1A and that RASSF1A reaches active Ras indirectly via RASSF5, positioning RASSF5 as the direct Ras-sensing member of a scaffold network.","evidence":"Reciprocal co-IP with domain-deletion constructs in COS cells","pmids":["11857081"],"confidence":"High","gaps":["Functional consequence of the heterodimer not resolved"]},{"year":2003,"claim":"Identified RASSF5/RAPL as a Rap1 effector controlling integrin function, extending its GTPase scaffolding role from apoptosis into lymphocyte adhesion and polarity.","evidence":"Co-IP, immunofluorescence, adhesion assays in lymphocytes after TCR/CXCL12 stimulation","pmids":["12845325"],"confidence":"High","gaps":["Did not identify the kinase partner mediating LFA-1 redistribution"]},{"year":2004,"claim":"Connected RASSF5 to MST1 and demonstrated bidirectional regulation—inhibiting autoactivation in solution but promoting activation at the membrane—revealing the Ras→RASSF5→MST apoptotic axis.","evidence":"Endogenous co-IP, in vitro kinase assay with purified proteins, membrane-targeting constructs","pmids":["15109305"],"confidence":"High","gaps":["Structural basis of homodimer disruption not yet defined","Physiological trigger for the inhibitor-to-activator switch unclear"]},{"year":2004,"claim":"Genetic loss-of-function in mice established RAPL as essential for chemokine-triggered lymphocyte and dendritic cell adhesion and in vivo trafficking.","evidence":"RAPL-knockout mouse, adhesion and in vivo trafficking assays","pmids":["15361866"],"confidence":"High","gaps":["Molecular link between Rap1, RAPL and LFA-1 redistribution not fully mapped"]},{"year":2004,"claim":"Showed RASSF5 growth suppression can be independent of its MST-, Ras-, and zinc-finger domains, implying additional uncharacterized effector routes.","evidence":"Colony formation and soft-agar assays with domain-deletion mutants","pmids":["15007383"],"confidence":"Medium","gaps":["The domain-independent effector was not identified","Single lab, no pathway mapping"]},{"year":2004,"claim":"Demonstrated Rap1-driven release of RAPL from microtubules to the leading edge, linking GTPase activation to cytoskeletal repositioning during directional migration.","evidence":"Live GFP imaging, FRET Rap1 probe, rap1GAPII expression, wound-healing assay in endothelial cells","pmids":["15569673"],"confidence":"Medium","gaps":["Mechanism of microtubule association not defined","Single lab"]},{"year":2006,"claim":"Established MST1 as the obligate kinase partner for RAPL-driven LFA-1 clustering, unifying the adhesion and Hippo-kinase functions of RASSF5.","evidence":"Co-IP, siRNA knockdown, co-localization, kinase and adhesion assays","pmids":["16892067"],"confidence":"High","gaps":["How MST1 kinase activity drives LFA-1 clustering mechanistically unresolved"]},{"year":2006,"claim":"Defined the autoinhibitory C1–RA intramolecular complex and its disruption by GTP-Ras, providing the structural switch that gates RASSF5 activity and lipid binding.","evidence":"NMR structure of C1 domain, ITC, chemical shift/relaxation measurements in mouse Nore1","pmids":["16698549"],"confidence":"High","gaps":["Did not directly link C1 release to SARAH/MST engagement","Membrane context not tested"]},{"year":2007,"claim":"Showed RASSF5 binds Rap2 and classical Ras more stably than Rap1 and that SARAH mediates its homodimerization, refining GTPase selectivity and the oligomeric basis of signaling.","evidence":"Co-IP, mutagenesis, 3D modeling, adhesion/migration assays","pmids":["17716979"],"confidence":"Medium","gaps":["Functional preference among GTPases in vivo not resolved","Single lab"]},{"year":2010,"claim":"Identified an MST-independent proliferation control: RAPL drives p27kip1 nuclear localization to restrain lymphocyte cycling, with loss causing autoimmunity and lymphoma.","evidence":"RAPL-knockout mouse, p27 Ser10 phospho-analysis, S10A knock-in rescue, Cdk2 assay","pmids":["21194982"],"confidence":"High","gaps":["How RAPL inhibits p27 Ser10 phosphorylation mechanistically unknown"]},{"year":2010,"claim":"Demonstrated RASSF5 mediates death-receptor apoptosis through MST1 and acts as a tumor suppressor restraining Ras transformation.","evidence":"siRNA knockdown, Rassf5-knockout mice and MEFs, apoptosis and transformation assays","pmids":["20810663"],"confidence":"High","gaps":["Connection between death-receptor engagement and RASSF5 activation undefined"]},{"year":2010,"claim":"Mapped NES/NLS signals and Lck-dependent phosphorylation controlling nucleocytoplasmic shuttling, tying subcellular localization to RASSF5's cell-cycle and apoptotic outputs.","evidence":"Heterokaryon shuttling, leptomycin B, mutagenesis, Lck co-IP, flow cytometry","pmids":["20064523"],"confidence":"Medium","gaps":["Nuclear substrates/targets of RASSF5 not identified","Single lab"]},{"year":2010,"claim":"Defined SKAP1 as a SARAH-binding partner required to deliver the Rap1–RapL–LFA-1 complex and modulate T cell motility, completing the adhesion-complex architecture.","evidence":"Skap1-knockout T cells, co-IP, L224A mutagenesis, intravital imaging","pmids":["20346707"],"confidence":"High","gaps":["Stoichiometry and dynamics of the full complex not resolved"]},{"year":2011,"claim":"Showed SKAP1's PH domain drives PI3K-dependent membrane recruitment of RapL, defining the membrane-targeting step that activates LFA-1.","evidence":"Co-IP, membrane fractionation, PH-domain mutation, myr-SKAP1 construct, LFA-1–ICAM-1 binding","pmids":["21669874"],"confidence":"Medium","gaps":["Interplay with the C1 lipid-binding interface not tested","Single lab"]},{"year":2012,"claim":"Revealed a scaffolding role beyond MST: RASSF5 bridges HIPK1 to Mdm2 to promote its ubiquitination, expanding its tumor-suppressive output.","evidence":"Endogenous co-IP, ubiquitination assay, Nore1-knockout MEFs, tumor assay","pmids":["22173032"],"confidence":"Medium","gaps":["Regulation of this scaffold function unknown","Single lab"]},{"year":2013,"claim":"Crystal structures of Mst2±RASSF5 SARAH explained the timing-dependent switch: RASSF5 disrupts the Mst2 activating homodimer only before activation-loop phosphorylation, otherwise leaving the active kinase intact.","evidence":"X-ray crystallography of Mst2 and Mst2–RASSF5 SARAH, kinase assay, mutagenesis","pmids":["23972470"],"confidence":"High","gaps":["What determines binding timing in cells not defined"]},{"year":2013,"claim":"Characterized the Nore1 SARAH homodimer as a lower-affinity antiparallel coiled coil relative to MST1, clarifying the thermodynamic basis for hetero- versus homo-dimer preference.","evidence":"X-ray crystallography, CD/DSF stability, analytical ultracentrifugation","pmids":["23331050"],"confidence":"High","gaps":["Cellular consequence of differential SARAH stability not tested"]},{"year":2013,"claim":"Identified acetylation-gated Itch-mediated ubiquitination as the route controlling RASSF5 protein stability and its apoptotic/cell-cycle activity.","evidence":"Co-IP, ubiquitination assay, acetylation analysis, ligase-dead control, flow cytometry","pmids":["23538446"],"confidence":"Medium","gaps":["Acetyltransferase/deacetylase governing the switch not identified","Single lab"]},{"year":2014,"claim":"Showed MST2 SARAH homodimerization and RAPL heterodimerization are both required for full kinase activation and T cell apoptosis, integrating structure with function.","evidence":"X-ray crystallography, structure-guided mutagenesis, kinase and apoptosis assays","pmids":["24468289"],"confidence":"High","gaps":["Order of homo- and hetero-dimerization events in vivo unresolved"]},{"year":2014,"claim":"Placed RASSF5 upstream of Ndr1/2 kinases in neuronal polarization via Par3 Ser383 phosphorylation, broadening its kinase-scaffold repertoire beyond MST.","evidence":"Neuronal shRNA, Ndr→Par3 phosphorylation assay, axon specification in hippocampal neurons","pmids":["24928906"],"confidence":"Medium","gaps":["How RASSF5 engages Ndr kinases mechanistically undefined","Single lab"]},{"year":2017,"claim":"Computational modeling proposed that the RA domain holds the SARAH domain in a kinked autoinhibited state relieved by K-Ras4B-GTP at the membrane, integrating the structural switch model.","evidence":"All-atom molecular dynamics simulations from crystal structures (no experimental validation)","pmids":["28197608"],"confidence":"Low","gaps":["Computational only; awaits experimental validation","Membrane requirement not tested biochemically"]},{"year":2019,"claim":"Established chromatin-level control of RASSF5 by Jmjd3-mediated H3K27me3 removal, linking epigenetic regulation to RASSF5-dependent apoptosis.","evidence":"ChIP for H3K27me3, Jmjd3 shRNA, qRT-PCR, apoptosis assays in osteoblasts","pmids":["31092054"],"confidence":"Medium","gaps":["Whether this regulation operates in cancer cells not tested","Single lab"]},{"year":2021,"claim":"Engineered high-affinity RA-domain variants showed that boosting Ras engagement simultaneously suppresses pro-cancer pathways and activates RASSF5 anti-cancer (senescence, p53) outputs, validating the effector axis as a therapeutic node.","evidence":"Computational design plus in vitro evolution, viability/migration/senescence assays, p53 modification blots in A549","pmids":["34717958"],"confidence":"Medium","gaps":["Mechanism linking RA binding to p53 modification not defined","Single lab"]},{"year":2023,"claim":"Defined an Irf8→RASSF5 transcriptional axis restraining emergency granulopoiesis, with loss driving clonal hematopoiesis and AML, extending its tumor-suppressor role to myeloid biology.","evidence":"Rassf5-knockout mouse, Irf8 ChIP/transcription assays, clonal hematopoiesis analysis","pmids":["37247756"],"confidence":"Medium","gaps":["Downstream effector mediating granulopoiesis control unknown","Single lab"]},{"year":null,"claim":"How the multiple inputs—GTPase loading, lipid binding, SKAP1/membrane recruitment, Lck phosphorylation, and acetylation/ubiquitination—are integrated to determine whether RASSF5 inhibits or activates MST kinases in a given cellular context remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unified model of context-dependent inhibitor-versus-activator switching","Endogenous timing of SARAH binding relative to MST activation in cells not measured","Domain-independent growth-suppressive effector still unidentified"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[3,7,13,15,16]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[3,16,19]},{"term_id":"GO:0008289","term_label":"lipid binding","supporting_discovery_ids":[8]}],"localization":[{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[6]},{"term_id":"GO:0031410","term_label":"cytoplasmic vesicle","supporting_discovery_ids":[7,13]},{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[3,14,21]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[12]}],"pathway":[{"term_id":"R-HSA-5357801","term_label":"Programmed Cell 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Seems to be involved in lymphocyte adhesion by linking RAP1A activation upon T-cell receptor or chemokine stimulation to integrin activation. Isoform 2 stimulates lymphocyte polarization and the patch-like distribution of ITGAL/LFA-1, resulting in an enhanced adhesion to ICAM1. Together with RAP1A may participate in regulation of microtubule growth. The association of isoform 2 with activated RAP1A is required for directional movement of endothelial cells during wound healing. May be involved in regulation of Ras apoptotic function. The RASSF5-STK4/MST1 complex may mediate HRAS and KRAS induced apoptosis","subcellular_location":"Cytoplasm; Cytoplasm, cytoskeleton","url":"https://www.uniprot.org/uniprotkb/Q8WWW0/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/RASSF5","classification":"Not Classified","n_dependent_lines":8,"n_total_lines":1208,"dependency_fraction":0.006622516556291391},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/RASSF5","total_profiled":1310},"omim":[{"mim_id":"609492","title":"RAS ASSOCIATION DOMAIN FAMILY PROTEIN 2; RASSF2","url":"https://www.omim.org/entry/609492"},{"mim_id":"607020","title":"RAS ASSOCIATION DOMAIN FAMILY PROTEIN 5; RASSF5","url":"https://www.omim.org/entry/607020"},{"mim_id":"600098","title":"RELATED RAS VIRAL ONCOGENE HOMOLOG 2; RRAS2","url":"https://www.omim.org/entry/600098"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"bone marrow","ntpm":103.2},{"tissue":"lymphoid 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RASSF1A association with RasG12V in COS cells is indirect, mediated by its heterodimerization with Nore1A via Nore1A's RA domain.\",\n      \"method\": \"Co-immunoprecipitation in COS cells; domain-deletion and co-expression experiments\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal co-IP with domain-deletion controls, multiple constructs tested, mechanistic conclusion supported by multiple orthogonal experiments\",\n      \"pmids\": [\"11857081\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"RASSF5/RAPL is a Rap1 effector that associates with active Rap1 after TCR and CXCL12 stimulation; it binds LFA-1 and mediates Rap1-triggered spatial redistribution of LFA-1 to the leading edge, stimulating lymphocyte polarization and adhesion to ICAM-1.\",\n      \"method\": \"Co-immunoprecipitation, immunofluorescence localization, overexpression in lymphocytes, adhesion assays\",\n      \"journal\": \"Nature immunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal co-IP, functional localization assays, adhesion readouts, replicated in primary and transfected cells\",\n      \"pmids\": [\"12845325\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"Endogenous MST1 immunoprecipitates with RASSF5/NORE1A from KB cells; recombinant NORE1A directly inhibits MST1 autoactivation in vitro by suppressing Thr183 autophosphorylation; membrane-targeted NORE1A or recruitment of MST1 to Ras(G12V) via NORE1A increases MST1 Thr183 phosphorylation.\",\n      \"method\": \"Endogenous co-IP, in vitro kinase assay with purified proteins, co-transfection with CAAX/myristoylation constructs\",\n      \"journal\": \"The Biochemical journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro reconstitution with purified proteins plus in-cell co-IP and multiple gain-of-function constructs\",\n      \"pmids\": [\"15109305\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"RASSF5/RAPL deficiency in mice causes defective chemokine-triggered lymphocyte adhesion and impaired migration to secondary lymphoid organs; dendritic cells from RAPL-deficient mice also show defective adhesion and fail to migrate to lymph nodes after inflammatory stimulation.\",\n      \"method\": \"RAPL-knockout mouse model, adhesion assays, in vivo trafficking experiments, histology\",\n      \"journal\": \"Nature immunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic knockout with defined cellular and in vivo phenotypes, multiple immune cell types examined\",\n      \"pmids\": [\"15361866\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"RASSF5/Nore1A and Nore1B suppress colony formation and anchorage-independent growth in specific tumor cell lines; growth inhibition by Nore1A is independent of its MST-binding domain, Ras-binding domain, and zinc-finger, indicating growth suppression through unidentified effectors.\",\n      \"method\": \"Colony formation assay, soft agar assay, domain-deletion mutants, cell cycle analysis\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function domain analysis in multiple cell lines, single lab, no pathway identification\",\n      \"pmids\": [\"15007383\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"RASSF5/RAPL localizes on microtubules in endothelial cells; activated Rap1 induces dissociation of RAPL from microtubules and its redistribution to the leading edge, controlling directional cell migration; disruption of Rap1-RAPL interaction perturbs wound healing.\",\n      \"method\": \"Fluorescence live imaging (GFP-RAPL), FRET-based Rap1 activation probe, adenoviral expression of rap1GAPII, wound healing assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — live imaging with functional consequence, FRET-based activity probe, single lab\",\n      \"pmids\": [\"15569673\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"RASSF5/RAPL binds and regulates the localization and kinase activity of Mst1; RAPL and Mst1 co-localize in vesicular compartments and translocate together with LFA-1 to the leading edge upon Rap1 activation; Mst1 knockdown abolishes RAPL-induced LFA-1 clustering, cell polarization, and adhesion.\",\n      \"method\": \"Co-immunoprecipitation, siRNA knockdown, immunofluorescence co-localization, kinase activity assay, adhesion assay\",\n      \"journal\": \"Nature immunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal co-IP, functional kinase assay, siRNA phenotype with specific readout, live localization, multiple orthogonal methods\",\n      \"pmids\": [\"16892067\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"In mouse Nore1, the C1 domain forms an intramolecular complex with the RA domain that weakens Ras/Rap1-GTP binding; GTP-Ras binding to the RA domain disrupts this C1-RA intramolecular complex, making the C1 domain's lipid-binding interface accessible; the free C1 domain binds phosphatidylinositol 3-phosphate.\",\n      \"method\": \"NMR structure of C1 domain, isothermal calorimetric titration (ITC), chemical shift and relaxation rate measurements\",\n      \"journal\": \"Structure\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — NMR structure with ITC binding measurements and chemical shift perturbation, single lab but multiple orthogonal structural/biophysical methods\",\n      \"pmids\": [\"16698549\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"RASSF5/RAPL forms more stable complexes with Rap2 and classical Ras than with Rap1; the SARAH domain mediates RAPL homodimerization in vitro and in cells; a single residue in switch I of Rap proteins (residue 39) critically determines the different interaction kinetics with RAPL; RAPL acts downstream of Rap2 in integrin-triggered T cell migration.\",\n      \"method\": \"Co-immunoprecipitation, mutagenesis, 3D modeling, adhesion and migration assays\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP with mutagenesis and functional assays, single lab, multiple constructs\",\n      \"pmids\": [\"17716979\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"RASSF5/RAPL suppresses lymphocyte proliferation by promoting nuclear localization of p27kip1: RAPL inhibits phosphorylation of p27kip1 on Ser10, thereby promoting its nuclear translocation; RAPL-deficient mice show cytoplasmic mislocalization of p27kip1, hyperproliferation, enhanced Cdk2 activity, and develop lupus-like disease and B cell lymphoma.\",\n      \"method\": \"RAPL-knockout mouse, flow cytometry (cell cycle), kinase assay (Cdk2), immunofluorescence localization of p27kip1, S10A knock-in rescue experiment\",\n      \"journal\": \"Immunity\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic knockout with in vivo phenotype, mechanistic rescue by S10A p27 mutation, multiple orthogonal methods\",\n      \"pmids\": [\"21194982\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"RASSF5/NORE1 mediates death receptor (TNF-α, TRAIL)-induced apoptosis through its interaction with MST1; siRNA depletion of RASSF5 reduces TNF-α-mediated apoptosis and Mst1 activation; Rassf5-null mice are resistant to TNF-α-induced apoptosis; Rassf5-null MEFs spontaneously immortalize and are transformable by K-RasG12V.\",\n      \"method\": \"siRNA knockdown, Rassf5-knockout mouse, MEF immortalization and transformation assays, apoptosis assays\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic knockout with in vivo apoptosis phenotype, siRNA knockdown in cell lines, MST1 activation assay, multiple orthogonal methods\",\n      \"pmids\": [\"20810663\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"RASSF5/RAPL contains functional nuclear export signal (NES, aa 260–300) dependent on CRM-1, and two nuclear localization signals; Lck tyrosine kinase binds RASSF5 through its C-terminal SH2-binding motif and phosphorylates it, which is required for efficient nuclear import of RASSF5; nuclear-localized (ΔNES) RASSF5 causes G1/S arrest and apoptosis, while an Lck-interaction-defective mutant induces apoptosis without cell cycle arrest.\",\n      \"method\": \"Heterokaryon shuttling assay, leptomycin B treatment, mutational analysis, immunofluorescence, co-immunoprecipitation with Lck, flow cytometry (cell cycle/apoptosis)\",\n      \"journal\": \"Journal of molecular biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP with mutagenesis, localization assays with functional consequences, single lab\",\n      \"pmids\": [\"20064523\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"SSKAP1 N-terminal domain binds the C-terminal SARAH domain of RASSF5/RapL; TCR-induced Rap1-RapL complex formation and LFA-1 binding fails in Skap1−/− T cells; a RapL mutation (L224A) abrogating SKAP1 binding disrupts colocalization in vesicles and T cell–DC conjugation; RapL expression slows T cell motility in lymph nodes.\",\n      \"method\": \"Skap1 knockout primary T cells, co-immunoprecipitation, point mutagenesis, intravital imaging of T cells in lymph nodes\",\n      \"journal\": \"Immunity\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic knockout, mutagenesis, co-IP, and intravital imaging with functional dwell-time readout\",\n      \"pmids\": [\"20346707\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"SKAP1 PH domain is required for RapL membrane translocation in a PI3K-dependent manner; membrane-targeted (myristoylated) SKAP1 constitutively recruits RapL to membranes and promotes Rap1 and LFA-1 binding, substituting for PI3K and TCR ligation in LFA-1 activation.\",\n      \"method\": \"Co-immunoprecipitation, membrane fractionation, PH-domain point mutation (R131M), myr-SKAP1 constitutive construct, LFA-1–ICAM-1 binding assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP with mutagenesis and gain-of-function constructs, single lab\",\n      \"pmids\": [\"21669874\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"RASSF5/NORE1 facilitates polyubiquitination and proteasomal degradation of oncoprotein HIPK1 by scaffolding its interaction with the Mdm2 E3 ubiquitin ligase; endogenous HIPK1 is stabilized in Nore1-deficient MEFs.\",\n      \"method\": \"Co-immunoprecipitation (endogenous), ubiquitination assay, Nore1-knockout MEFs, in vivo tumor formation assay\",\n      \"journal\": \"EMBO reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP plus genetic KO validation, single lab, ubiquitination assay\",\n      \"pmids\": [\"22173032\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"Crystal structures of human Mst2 alone and in complex with RASSF5 SARAH domain reveal that: (i) Mst2 activates by transautophosphorylation at its activation loop requiring SARAH-mediated homodimerization; (ii) RASSF5 SARAH disrupts the Mst2 homodimer and blocks Mst2 autoactivation when bound before activation-loop phosphorylation; (iii) RASSF5 binding to already-activated Mst2 does not inhibit kinase activity—suggesting RASSF5 can be either inhibitor or positive regulator depending on the timing of binding relative to Mst2 activation.\",\n      \"method\": \"X-ray crystallography (Mst2 alone and Mst2–RASSF5 SARAH complex), in vitro kinase assay, mutagenesis\",\n      \"journal\": \"Structure\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structures with in vitro kinase assay and mutagenesis, multiple orthogonal methods in single rigorous study\",\n      \"pmids\": [\"23972470\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"The SARAH domain of RASSF5/Nore1 crystallizes as an antiparallel homodimeric coiled coil with heptad register interrupted by two stutters; the Nore1 SARAH homodimer has lower affinity and thermodynamic stability than the MST1 SARAH homodimer; the SARAH domain undergoes association-dependent folding.\",\n      \"method\": \"X-ray crystallography of SARAH homodimer, thermodynamic stability measurements (CD, DSF), analytical ultracentrifugation\",\n      \"journal\": \"Biochemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — crystal structure with thermodynamic characterization, single lab but multiple biophysical methods\",\n      \"pmids\": [\"23331050\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"E3 ubiquitin ligase Itch binds RASSF5 through its WW domains interacting with the PPxY motif of RASSF5; Itch overexpression induces RASSF5 poly-ubiquitination and proteasomal degradation; acetylation of RASSF5 in tumor cells blocks Itch binding, stabilizing RASSF5; inhibition of RASSF5 acetylation restores Itch binding and triggers degradation; Itch overexpression (but not ligase-dead mutant) abrogates RASSF5-mediated G1 arrest and apoptosis.\",\n      \"method\": \"Co-immunoprecipitation (in vivo and in vitro), ubiquitination assay, flow cytometry (cell cycle/apoptosis), acetylation analysis\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP in vivo and in vitro, ubiquitination assay, ligase-dead control, single lab\",\n      \"pmids\": [\"23538446\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Crystal structure of MST2 SARAH domain forms antiparallel homodimeric coiled coil; residues critical for MST2 homodimerization also impair its heterodimerization with RASSF5/RAPL SARAH domain; SARAH-mediated homodimerization and heterodimerization with RAPL are both required for full MST2 kinase activation and apoptosis in T cells.\",\n      \"method\": \"X-ray crystallography, structure-guided mutagenesis, kinase activity assay, apoptosis assay in T cells\",\n      \"journal\": \"Journal of structural biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — crystal structure plus mutagenesis with functional kinase and apoptosis assays, single lab\",\n      \"pmids\": [\"24468289\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"RASSF5 acts upstream of Ndr1/Ndr2 kinases in a novel signaling cascade in neurons; Rassf5 and Ndr1/2 are required for hippocampal neuron polarization; Ndr kinases phosphorylate Par3 at Ser383 to inhibit Par3–dynein interaction, thereby polarizing Par3 distribution and specifying a single axon.\",\n      \"method\": \"Neuronal knockdown (shRNA), phosphorylation assay (Ndr→Par3 Ser383), Par3 localization by immunofluorescence, axon specification assay in primary hippocampal neurons\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — shRNA knockdown with defined substrate phosphorylation and rescue, single lab\",\n      \"pmids\": [\"24928906\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"In inactive RASSF5, the RA domain retains the SARAH domain in a self-associated autoinhibited conformation (kinked α-helix); K-Ras4B-GTP binding shifts the equilibrium toward SARAH domain interaction with MST kinases, enabling SARAH heterodimerization and MST1/2 kinase domain trans-autophosphorylation; membrane context is required for productive MST activation.\",\n      \"method\": \"All-atom molecular dynamics simulations based on crystal structures of RA and SARAH domains\",\n      \"journal\": \"Physical chemistry chemical physics\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 4 / Weak — computational modeling only, no experimental validation in this paper\",\n      \"pmids\": [\"28197608\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Histone demethylase Jmjd3 (KDM6B) directly targets the RASSF5 promoter; Jmjd3 knockdown increases H3K27me3 at the RASSF5 promoter and decreases RASSF5 expression, reducing TNF-α-induced osteoblast apoptosis; Jmjd3 thus regulates apoptosis through RASSF5.\",\n      \"method\": \"ChIP assay (H3K27me3 at RASSF5 promoter), shRNA knockdown of Jmjd3, qRT-PCR, apoptosis assays (Annexin V, caspase-3)\",\n      \"journal\": \"Connective tissue research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP with functional apoptosis assay, single lab\",\n      \"pmids\": [\"31092054\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Engineered high-affinity RASSF5 RA-domain variants, generated by computational design and in vitro evolution, inhibit Ras-regulated pro-cancer pathways and simultaneously stimulate RASSF5 anti-cancer pathways; introduction into A549 cells decreases viability and motility and induces cellular senescence with increased p53 acetylation and decreased p53 phosphorylation, to a greater extent than WT RASSF5.\",\n      \"method\": \"Computational design plus in vitro evolution (phage/yeast display implied), cell viability assay, migration assay, senescence assay, p53 modification Western blot in A549 cells\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — protein engineering with multiple cellular functional readouts, single lab\",\n      \"pmids\": [\"34717958\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Irf8 transcriptionally activates RASSF5 (Nore1a) expression during emergency granulopoiesis; Rassf5-knockout mice develop neutrophilia and progress to AML with aging, display sustained emergency granulopoiesis, and show enhanced DNA damage and clonal hematopoiesis in hematopoietic stem cells.\",\n      \"method\": \"Rassf5-knockout mouse model, gene expression analysis, clonal hematopoiesis analysis, Irf8 ChIP/transcription assays\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic knockout with in vivo phenotype, Irf8 target gene validation, single lab\",\n      \"pmids\": [\"37247756\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"RASSF5 (NORE1/RAPL) is a non-catalytic scaffold protein that binds GTP-loaded Ras and Rap1 GTPases via its RA domain and interacts with the pro-apoptotic Ste20-like kinases MST1/2 via SARAH domain heterodimerization; in the autoinhibited state its C1 domain occludes the RA domain, but GTP-Ras binding relieves this and can drive membrane recruitment, where RASSF5 disrupts MST2 homodimers to block autoactivation before MST2 phosphorylation yet can positively regulate already-activated MST2 — together these activities link Ras signaling to the Hippo pathway to promote apoptosis and suppress proliferation; in lymphocytes RASSF5/RAPL forms a Rap1-SKAP1-RapL-LFA-1 complex that, together with MST1, spatially redistributes LFA-1 to the leading edge to drive adhesion and cell polarity, and additionally controls lymphocyte cell cycle entry by promoting nuclear localization of p27kip1; RASSF5 protein stability is regulated by acetylation-gated ubiquitination by Itch and by Lck-mediated tyrosine phosphorylation that controls its nucleocytoplasmic shuttling, while at the transcriptional level it is epigenetically silenced by promoter methylation in many cancers and activated by TET1-mediated demethylation and Jmjd3-mediated H3K27me3 removal.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"RASSF5 (NORE1/RAPL) is a non-catalytic scaffold that couples Ras-family GTPase signaling to the pro-apoptotic Hippo-related kinases MST1/2, linking growth-factor and adhesion cues to apoptosis, cell-cycle control, and cytoskeletal polarity [#0, #3, #11]. It binds GTP-loaded Ras through its RA domain following EGF receptor activation [#0] and engages MST1/2 through SARAH-domain heterodimerization [#3, #16]. Structural work resolves the regulatory logic: in the inactive state an intramolecular C1\\u2013RA complex occludes the RA domain and renders the SARAH domain self-associated and autoinhibited, while GTP-Ras binding disrupts the C1\\u2013RA complex, exposing a phosphatidylinositol-3-phosphate-binding C1 interface and freeing SARAH to heterodimerize with MST kinases [#8, #16]. RASSF5 acts as a timing-dependent switch on MST2\\u2014its SARAH domain disrupts the MST2 homodimer to block trans-autophosphorylation when bound before activation, yet does not inhibit already-activated kinase, allowing it to be either inhibitor or positive regulator [#16, #3]. Through MST1, RASSF5 mediates death-receptor (TNF-\\u03b1/TRAIL)-induced apoptosis and suppresses transformation, with Rassf5-null cells immortalizing and becoming transformable by K-RasG12V [#11]. As the Rap1/Rap2 effector RAPL, it scaffolds a Rap1\\u2013SKAP1\\u2013RAPL\\u2013LFA-1 complex that, with MST1, spatially redistributes LFA-1 to the leading edge to drive lymphocyte adhesion, polarity, and trafficking [#2, #7, #13], and it independently restrains lymphocyte proliferation by promoting nuclear localization of p27kip1 [#10]. RASSF5 stability and localization are tightly controlled: acetylation-gated, Itch-mediated ubiquitination governs its turnover [#18], and Lck-dependent tyrosine phosphorylation controls its CRM1-dependent nucleocytoplasmic shuttling and pro-apoptotic activity [#12]. At the transcriptional level its expression is set by chromatin regulators including Jmjd3-mediated H3K27me3 removal and Irf8-driven activation during granulopoiesis [#22, #24].\",\n  \"teleology\": [\n    {\n      \"year\": 1998,\n      \"claim\": \"Established RASSF5 as a candidate Ras effector by showing direct GTP-dependent Ras binding, defining the entry point linking it to Ras signaling.\",\n      \"evidence\": \"GTP-dependent in vitro binding assay and co-IP after EGF stimulation in COS-7 and KB cells\",\n      \"pmids\": [\"9488663\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not define a downstream effector function\", \"No structural basis for binding\"]\n    },\n    {\n      \"year\": 2002,\n      \"claim\": \"Showed RASSF5 hetero-oligomerizes with RASSF1A and that RASSF1A reaches active Ras indirectly via RASSF5, positioning RASSF5 as the direct Ras-sensing member of a scaffold network.\",\n      \"evidence\": \"Reciprocal co-IP with domain-deletion constructs in COS cells\",\n      \"pmids\": [\"11857081\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional consequence of the heterodimer not resolved\"]\n    },\n    {\n      \"year\": 2003,\n      \"claim\": \"Identified RASSF5/RAPL as a Rap1 effector controlling integrin function, extending its GTPase scaffolding role from apoptosis into lymphocyte adhesion and polarity.\",\n      \"evidence\": \"Co-IP, immunofluorescence, adhesion assays in lymphocytes after TCR/CXCL12 stimulation\",\n      \"pmids\": [\"12845325\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not identify the kinase partner mediating LFA-1 redistribution\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Connected RASSF5 to MST1 and demonstrated bidirectional regulation\\u2014inhibiting autoactivation in solution but promoting activation at the membrane\\u2014revealing the Ras\\u2192RASSF5\\u2192MST apoptotic axis.\",\n      \"evidence\": \"Endogenous co-IP, in vitro kinase assay with purified proteins, membrane-targeting constructs\",\n      \"pmids\": [\"15109305\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of homodimer disruption not yet defined\", \"Physiological trigger for the inhibitor-to-activator switch unclear\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Genetic loss-of-function in mice established RAPL as essential for chemokine-triggered lymphocyte and dendritic cell adhesion and in vivo trafficking.\",\n      \"evidence\": \"RAPL-knockout mouse, adhesion and in vivo trafficking assays\",\n      \"pmids\": [\"15361866\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular link between Rap1, RAPL and LFA-1 redistribution not fully mapped\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Showed RASSF5 growth suppression can be independent of its MST-, Ras-, and zinc-finger domains, implying additional uncharacterized effector routes.\",\n      \"evidence\": \"Colony formation and soft-agar assays with domain-deletion mutants\",\n      \"pmids\": [\"15007383\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"The domain-independent effector was not identified\", \"Single lab, no pathway mapping\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Demonstrated Rap1-driven release of RAPL from microtubules to the leading edge, linking GTPase activation to cytoskeletal repositioning during directional migration.\",\n      \"evidence\": \"Live GFP imaging, FRET Rap1 probe, rap1GAPII expression, wound-healing assay in endothelial cells\",\n      \"pmids\": [\"15569673\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism of microtubule association not defined\", \"Single lab\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Established MST1 as the obligate kinase partner for RAPL-driven LFA-1 clustering, unifying the adhesion and Hippo-kinase functions of RASSF5.\",\n      \"evidence\": \"Co-IP, siRNA knockdown, co-localization, kinase and adhesion assays\",\n      \"pmids\": [\"16892067\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How MST1 kinase activity drives LFA-1 clustering mechanistically unresolved\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Defined the autoinhibitory C1\\u2013RA intramolecular complex and its disruption by GTP-Ras, providing the structural switch that gates RASSF5 activity and lipid binding.\",\n      \"evidence\": \"NMR structure of C1 domain, ITC, chemical shift/relaxation measurements in mouse Nore1\",\n      \"pmids\": [\"16698549\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not directly link C1 release to SARAH/MST engagement\", \"Membrane context not tested\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Showed RASSF5 binds Rap2 and classical Ras more stably than Rap1 and that SARAH mediates its homodimerization, refining GTPase selectivity and the oligomeric basis of signaling.\",\n      \"evidence\": \"Co-IP, mutagenesis, 3D modeling, adhesion/migration assays\",\n      \"pmids\": [\"17716979\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional preference among GTPases in vivo not resolved\", \"Single lab\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Identified an MST-independent proliferation control: RAPL drives p27kip1 nuclear localization to restrain lymphocyte cycling, with loss causing autoimmunity and lymphoma.\",\n      \"evidence\": \"RAPL-knockout mouse, p27 Ser10 phospho-analysis, S10A knock-in rescue, Cdk2 assay\",\n      \"pmids\": [\"21194982\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How RAPL inhibits p27 Ser10 phosphorylation mechanistically unknown\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Demonstrated RASSF5 mediates death-receptor apoptosis through MST1 and acts as a tumor suppressor restraining Ras transformation.\",\n      \"evidence\": \"siRNA knockdown, Rassf5-knockout mice and MEFs, apoptosis and transformation assays\",\n      \"pmids\": [\"20810663\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Connection between death-receptor engagement and RASSF5 activation undefined\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Mapped NES/NLS signals and Lck-dependent phosphorylation controlling nucleocytoplasmic shuttling, tying subcellular localization to RASSF5's cell-cycle and apoptotic outputs.\",\n      \"evidence\": \"Heterokaryon shuttling, leptomycin B, mutagenesis, Lck co-IP, flow cytometry\",\n      \"pmids\": [\"20064523\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Nuclear substrates/targets of RASSF5 not identified\", \"Single lab\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Defined SKAP1 as a SARAH-binding partner required to deliver the Rap1\\u2013RapL\\u2013LFA-1 complex and modulate T cell motility, completing the adhesion-complex architecture.\",\n      \"evidence\": \"Skap1-knockout T cells, co-IP, L224A mutagenesis, intravital imaging\",\n      \"pmids\": [\"20346707\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Stoichiometry and dynamics of the full complex not resolved\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Showed SKAP1's PH domain drives PI3K-dependent membrane recruitment of RapL, defining the membrane-targeting step that activates LFA-1.\",\n      \"evidence\": \"Co-IP, membrane fractionation, PH-domain mutation, myr-SKAP1 construct, LFA-1\\u2013ICAM-1 binding\",\n      \"pmids\": [\"21669874\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Interplay with the C1 lipid-binding interface not tested\", \"Single lab\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Revealed a scaffolding role beyond MST: RASSF5 bridges HIPK1 to Mdm2 to promote its ubiquitination, expanding its tumor-suppressive output.\",\n      \"evidence\": \"Endogenous co-IP, ubiquitination assay, Nore1-knockout MEFs, tumor assay\",\n      \"pmids\": [\"22173032\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Regulation of this scaffold function unknown\", \"Single lab\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Crystal structures of Mst2\\u00b1RASSF5 SARAH explained the timing-dependent switch: RASSF5 disrupts the Mst2 activating homodimer only before activation-loop phosphorylation, otherwise leaving the active kinase intact.\",\n      \"evidence\": \"X-ray crystallography of Mst2 and Mst2\\u2013RASSF5 SARAH, kinase assay, mutagenesis\",\n      \"pmids\": [\"23972470\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"What determines binding timing in cells not defined\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Characterized the Nore1 SARAH homodimer as a lower-affinity antiparallel coiled coil relative to MST1, clarifying the thermodynamic basis for hetero- versus homo-dimer preference.\",\n      \"evidence\": \"X-ray crystallography, CD/DSF stability, analytical ultracentrifugation\",\n      \"pmids\": [\"23331050\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Cellular consequence of differential SARAH stability not tested\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Identified acetylation-gated Itch-mediated ubiquitination as the route controlling RASSF5 protein stability and its apoptotic/cell-cycle activity.\",\n      \"evidence\": \"Co-IP, ubiquitination assay, acetylation analysis, ligase-dead control, flow cytometry\",\n      \"pmids\": [\"23538446\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Acetyltransferase/deacetylase governing the switch not identified\", \"Single lab\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Showed MST2 SARAH homodimerization and RAPL heterodimerization are both required for full kinase activation and T cell apoptosis, integrating structure with function.\",\n      \"evidence\": \"X-ray crystallography, structure-guided mutagenesis, kinase and apoptosis assays\",\n      \"pmids\": [\"24468289\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Order of homo- and hetero-dimerization events in vivo unresolved\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Placed RASSF5 upstream of Ndr1/2 kinases in neuronal polarization via Par3 Ser383 phosphorylation, broadening its kinase-scaffold repertoire beyond MST.\",\n      \"evidence\": \"Neuronal shRNA, Ndr\\u2192Par3 phosphorylation assay, axon specification in hippocampal neurons\",\n      \"pmids\": [\"24928906\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"How RASSF5 engages Ndr kinases mechanistically undefined\", \"Single lab\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Computational modeling proposed that the RA domain holds the SARAH domain in a kinked autoinhibited state relieved by K-Ras4B-GTP at the membrane, integrating the structural switch model.\",\n      \"evidence\": \"All-atom molecular dynamics simulations from crystal structures (no experimental validation)\",\n      \"pmids\": [\"28197608\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Computational only; awaits experimental validation\", \"Membrane requirement not tested biochemically\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Established chromatin-level control of RASSF5 by Jmjd3-mediated H3K27me3 removal, linking epigenetic regulation to RASSF5-dependent apoptosis.\",\n      \"evidence\": \"ChIP for H3K27me3, Jmjd3 shRNA, qRT-PCR, apoptosis assays in osteoblasts\",\n      \"pmids\": [\"31092054\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether this regulation operates in cancer cells not tested\", \"Single lab\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Engineered high-affinity RA-domain variants showed that boosting Ras engagement simultaneously suppresses pro-cancer pathways and activates RASSF5 anti-cancer (senescence, p53) outputs, validating the effector axis as a therapeutic node.\",\n      \"evidence\": \"Computational design plus in vitro evolution, viability/migration/senescence assays, p53 modification blots in A549\",\n      \"pmids\": [\"34717958\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism linking RA binding to p53 modification not defined\", \"Single lab\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Defined an Irf8\\u2192RASSF5 transcriptional axis restraining emergency granulopoiesis, with loss driving clonal hematopoiesis and AML, extending its tumor-suppressor role to myeloid biology.\",\n      \"evidence\": \"Rassf5-knockout mouse, Irf8 ChIP/transcription assays, clonal hematopoiesis analysis\",\n      \"pmids\": [\"37247756\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Downstream effector mediating granulopoiesis control unknown\", \"Single lab\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How the multiple inputs\\u2014GTPase loading, lipid binding, SKAP1/membrane recruitment, Lck phosphorylation, and acetylation/ubiquitination\\u2014are integrated to determine whether RASSF5 inhibits or activates MST kinases in a given cellular context remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No unified model of context-dependent inhibitor-versus-activator switching\", \"Endogenous timing of SARAH binding relative to MST activation in cells not measured\", \"Domain-independent growth-suppressive effector still unidentified\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [3, 7, 13, 15, 16]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [3, 16, 19]},\n      {\"term_id\": \"GO:0008289\", \"supporting_discovery_ids\": [8]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [6]},\n      {\"term_id\": \"GO:0031410\", \"supporting_discovery_ids\": [7, 13]},\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [3, 14, 21]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [12]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [11, 16, 19]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 3, 11]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [2, 4, 13, 24]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [10, 12]}\n    ],\n    \"complexes\": [\"Rap1-SKAP1-RapL-LFA-1 complex\", \"RASSF5-MST1/2 SARAH heterodimer\"],\n    \"partners\": [\"HRAS\", \"RAP1\", \"MST1\", \"STK4\", \"STK3\", \"SKAP1\", \"ITCH\", \"LCK\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":8,"faith_total":8,"faith_pct":100.0}}