{"gene":"MISP","run_date":"2026-06-10T02:59:50","timeline":{"discoveries":[{"year":2013,"finding":"MISP (C19orf21) is a substrate of Plk1; Plk1 phosphorylates MISP, and this phosphorylation is required for the metaphase-to-anaphase transition and proper mitotic spindle positioning. MISP is an actin-associated protein that forms a complex with p150(glued) (a dynein-dynactin subunit) and regulates its cortical distribution, thereby stabilizing cortical and astral microtubule attachments.","method":"Loss-of-function (siRNA depletion), phosphorylation assays identifying Plk1 as the kinase, co-immunoprecipitation of MISP with p150(glued), live-cell imaging of spindle orientation and astral microtubule length","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, kinase substrate assay, KD with defined spindle phenotype; replicated independently in a second concurrent study (PMID:23574715)","pmids":["23509069"],"is_preprint":false},{"year":2013,"finding":"MISP associates with the actin cytoskeleton and focal adhesions and is expressed only in adherent cell types. During mitosis, MISP is phosphorylated by Cdk1 and localizes to retraction fibers. MISP interacts with the +TIP protein EB1 and with p150(glued) (dynein/dynactin). Depletion of MISP causes mitotic arrest with reduced tension across sister kinetochores, chromosome misalignment, spindle multipolarity (in cells with supernumerary centrosomes), and randomized spindle orientation.","method":"Co-immunoprecipitation (MISP with EB1 and p150glued), Cdk1 phosphorylation assay, siRNA depletion with live-cell imaging, immunofluorescence of focal adhesions and retraction fibers, kinetochore tension measurements","journal":"Cell cycle (Georgetown, Tex.)","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, kinase assay, KD with multiple orthogonal phenotypic readouts; independent replication of core findings (PMID:23509069)","pmids":["23574715"],"is_preprint":false},{"year":2014,"finding":"MISP/Caprice is a direct actin-binding protein with multiple actin-binding sites that forms mesh-like F-actin bundles in vitro. Overexpression induces stress fiber-like thick filaments and knockdown causes filopodial formation, demonstrating a direct role in actin cytoskeletal organization. It is expressed in cells/tissues with specialized actin structures such as growth cones and inner ear hair cell stereocilia.","method":"In vitro F-actin bundling assay with purified protein, overexpression and knockdown in cultured cells with morphological readout, fractionation identifying MISP in highly insoluble scaffold fraction","journal":"Genes to cells : devoted to molecular & cellular mechanisms","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro bundling assay with purified protein plus loss- and gain-of-function cellular phenotypes in a single study","pmids":["24475924"],"is_preprint":false},{"year":2018,"finding":"MISP functions downstream of the ERM family member ezrin and upstream of NuMA in the spindle orientation pathway. SLK/LOK-activated (phosphorylated) ezrin competes with MISP for actin-binding sites at the cell cortex, limiting cortical MISP levels. Excessive MISP accumulation disrupts crescent-like NuMA polarization at the cortex and reduces astral microtubule dynamics. MISP directly interacts with ezrin.","method":"Co-immunoprecipitation (MISP–ezrin interaction), siRNA epistasis experiments (SLK/LOK, ezrin, MISP), live-cell imaging of NuMA localization and astral microtubule dynamics, cortical fractionation","journal":"Journal of cell science","confidence":"High","confidence_rationale":"Tier 2 / Moderate — direct Co-IP, genetic epistasis with defined pathway ordering, multiple orthogonal phenotypic readouts in a single study","pmids":["29669740"],"is_preprint":false},{"year":2018,"finding":"MISP interacts with the scaffolding protein IQGAP1, and through IQGAP1 binds the active (GTP-bound) form of Cdc42. MISP depletion increases IQGAP1 cortical accumulation and decreases Cdc42 activity. IQGAP1 overexpression rescues MISP-depletion phenotypes (spindle misorientation, loss of astral microtubules, prolonged mitosis) and restores active Cdc42 levels, placing IQGAP1 downstream of MISP. IQGAP1 also acts downstream of MISP in controlling p150(glued) localization.","method":"Co-immunoprecipitation (MISP–IQGAP1, IQGAP1–Cdc42), siRNA depletion epistasis, active-Cdc42 pull-down assay, IQGAP1 overexpression rescue experiments, immunofluorescence of p150(glued) localization","journal":"Scientific reports","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal Co-IP, active-Cdc42 pull-down, rescue epistasis, multiple orthogonal phenotypic readouts in one study","pmids":["29679050"],"is_preprint":false},{"year":2021,"finding":"Phosphorylation of MISP at S394, S395, and S400 increases actin-bundling activity (but not actin-binding activity), inducing stress fiber formation in interphase cells. Mitotic phosphorylation at S376, S471, and S541 suppresses actin-binding activity. Thus, cell-cycle-dependent phosphorylation differentially regulates MISP actin-binding versus bundling activities.","method":"Site-directed mutagenesis of phosphorylation sites, in vitro actin-binding and bundling assays, overexpression of phosphomutants in cells with morphological readout","journal":"Biochemical and biophysical research communications","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro biochemical assays combined with mutagenesis and cellular phenotype in a single study","pmids":["34023777"],"is_preprint":false},{"year":2022,"finding":"MISP localizes specifically to the rootlet end (basal, pointed-end side) of microvillar actin core bundles in transporting epithelia. Purified MISP exhibits potent F-actin bundling activity in vitro and promotes rootlet elongation in cells. MISP-bundled filaments recruit fimbrin, which further elongates and stabilizes bundles. Ezrin prevents MISP from decorating the membrane-wrapped distal end of the core bundle, confining MISP to the rootlet.","method":"Immunofluorescence/live imaging for localization, in vitro actin bundling assay with purified protein, siRNA/CRISPR KO with microvillar phenotype readout (rootlet length), co-sedimentation assays, ezrin loss-of-function experiments","journal":"Cell reports","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro reconstitution with purified protein, cellular loss-of-function with defined morphological phenotype, mechanistic dissection of ezrin competition; multiple orthogonal methods","pmids":["35443169"],"is_preprint":false},{"year":2023,"finding":"MISP downregulation by M2 macrophage-derived extracellular vesicles promotes IQGAP1 nuclear translocation and activates STAT3 phosphorylation in hepatocellular carcinoma cells, leading to upregulation of PD-L1 and immune escape. MISP, IQGAP1, and STAT3 interact in a complex as shown by co-immunoprecipitation. MISP negatively correlates with IQGAP1/PD-L1 expression.","method":"Co-immunoprecipitation (MISP–IQGAP1–STAT3 complex), MISP/IQGAP1 overexpression experiments, flow cytometry of CD8+ T cells, ELISA for cytokines, in vivo nude mouse xenograft","journal":"International immunopharmacology","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — Co-IP complex validated, in vivo rescue experiment, but single lab with limited mechanistic depth on MISP's direct biochemical role","pmids":["37633233"],"is_preprint":false},{"year":2023,"finding":"In intrahepatic cholangiocarcinoma cells, MISP localizes to adherens junctions and suppresses E-cadherin dimerization. PLK1 phosphorylates MISP (confirmed as a PLK1 substrate in this context) and, together with MISP, disrupts adherens junctions to promote lymphatic invasion and cell motility.","method":"siRNA knockdown of PLK1 or MISP with trans-lymphatic endothelial migration and wound healing assays, immunofluorescence of focal adhesions and adherens junctions, in vivo tumorigenesis assay","journal":"Cancer gene therapy","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — KD with defined cellular phenotypes, localization by IF, single lab without in vitro reconstitution of E-cadherin dimerization suppression","pmids":["38057358"],"is_preprint":false},{"year":2024,"finding":"MISP directly binds the SARAH domain of MST1/2 kinases, inhibiting their homodimerization and autophosphorylation, thereby sustaining YAP activation (Hippo pathway inhibition). YAP activation in turn increases SLC7A11 expression to protect cells from ferroptosis. A MISP-R390/391A mutant that disrupts MISP–MST1/2 binding abolishes this effect. YAP also transcriptionally activates MISP, creating a positive feedback loop.","method":"Co-immunoprecipitation (MISP–MST1/2), site-directed mutagenesis (R390/391A), in vitro kinase/autophosphorylation assay, siRNA knockdown with ferroptosis sensitivity readout, Western blot for Hippo pathway components, in vivo xenograft","journal":"Advanced science (Weinheim, Baden-Wurttemberg, Germany)","confidence":"High","confidence_rationale":"Tier 1 / Moderate — Co-IP, mutagenesis disrupting binding, in vitro kinase assay, multiple orthogonal readouts in one study with in vivo validation","pmids":["40019375"],"is_preprint":false},{"year":2024,"finding":"MISP preferentially binds ADP-actin (aged/pointed-end-enriched) filaments over ADP-Pi-actin filaments, and in vitro TIRF assays show MISP binds at or near the pointed ends of growing actin filaments. In solution, MISP assembles parallel bundles of uniform polarity, whereas substrate-attached MISP can bundle filaments in both parallel and antiparallel configurations. Microvillar rootlets are decorated with cofilin (indicating high ADP-actin content), consistent with MISP's nucleotide-state preference driving its rootlet localization.","method":"In vitro TIRF microscopy with purified MISP and differentially nucleotide-loaded actin filaments, immunostaining of native intestinal tissue for cofilin, in vitro bundling assays in solution vs. substrate-attached conditions","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro reconstitution with purified protein under multiple controlled conditions, corroborated by native tissue immunostaining; also supported by companion preprint (PMID:37205433)","pmids":["38588808","37205433"],"is_preprint":false},{"year":2024,"finding":"MISP promotes nuclear accumulation of β-catenin, thereby activating the Wnt/β-catenin signaling pathway in pancreatic cancer cells. Fisetin suppresses MISP expression by downregulating the transcription factor MYB, identifying MYB as a transcriptional activator of MISP.","method":"Overexpression/knockdown of MISP with Western blot for β-catenin nuclear localization, in vitro proliferation assays, in vivo xenograft, drug screening library","journal":"Biochimica et biophysica acta. Molecular basis of disease","confidence":"Medium","confidence_rationale":"Tier 3 / Weak — cellular overexpression/KD with β-catenin localization readout, single lab without direct biochemical reconstitution of the MISP–β-catenin interaction","pmids":["39278512"],"is_preprint":false},{"year":2024,"finding":"MISP forms a complex with Opa Interacting Protein 5 (OIP5) in the cytoplasm, and MISP influences OIP5 expression in a unidirectional manner. MISP also increases phosphorylated STAT3 levels via the JAK2-STAT3 signaling pathway in colorectal cancer cells.","method":"Co-immunoprecipitation (MISP–OIP5 complex), Western blot for pSTAT3, MISP-deficient mouse model (AOM/DSS colitis-induced tumors), siRNA knockdown with cell proliferation readout","journal":"International journal of molecular sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — Co-IP for complex, in vivo KO model with tumor phenotype, but limited mechanistic depth on how MISP activates JAK2-STAT3; single lab","pmids":["38474305"],"is_preprint":false},{"year":2025,"finding":"MISP directly interacts with MST1 (co-immunoprecipitation confirmed), inhibiting Hippo signaling and leading to increased YAP, TAZ, and CYR61 expression and decreased MST1 and phospho-YAP levels in lung adenocarcinoma cells.","method":"Co-immunoprecipitation (MISP–MST1), Western blot for Hippo pathway components, overexpression/knockdown with in vitro and in vivo proliferation and migration assays","journal":"Discover oncology","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — Co-IP confirming direct interaction with MST1, cellular phenotype with pathway readout, but single lab and limited mechanistic depth; largely corroborates PMID:40019375","pmids":["41249743"],"is_preprint":false},{"year":2022,"finding":"MISP-deficient mice show exacerbated DSS-induced colitis, with decreased intestinal epithelial cell proliferation and significantly reduced Tgfb1 (TGF-β1) anti-inflammatory cytokine expression in the colon, indicating MISP supports colon recovery from inflammation partly through TGF-β1 signaling.","method":"MISP-knockout mouse model (DSS colitis), RT-qPCR for Tgfb1 and other cytokines, histological assessment of crypt morphology, Ki67 proliferation staining","journal":"The Journal of veterinary medical science","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — defined KO model with specific molecular readout (Tgfb1), single lab without mechanistic reconstitution of how MISP regulates TGF-β1","pmids":["36596561"],"is_preprint":false}],"current_model":"MISP (C19orf21/Caprice) is a cell-cycle-regulated actin-binding and F-actin bundling protein that, during mitosis, is phosphorylated by Plk1 and Cdk1 to localize to the cell cortex and retraction fibers where it interacts with EB1, p150(glued)/dynactin, and IQGAP1–Cdc42 to stabilize astral microtubule attachments and orient the mitotic spindle; in interphase epithelial cells, MISP preferentially binds ADP-actin at filament pointed ends to bundle rootlet actin in microvilli, a localization enforced by competition with ezrin at the distal core bundle; beyond cytoskeletal roles, MISP inhibits MST1/2 homodimerization and autophosphorylation to suppress Hippo signaling and sustain YAP activation (which in turn transcriptionally upregulates MISP), and MISP additionally modulates IQGAP1/STAT3/PD-L1 and Wnt/β-catenin pathways in cancer contexts."},"narrative":{"mechanistic_narrative":"MISP (C19orf21/Caprice) is a cell-cycle-regulated, direct actin-binding and F-actin-bundling protein that couples cortical actin organization to mitotic spindle positioning and, in epithelia, to microvillar architecture [PMID:23509069, PMID:24475924, PMID:35443169]. During mitosis it is phosphorylated by Plk1 and Cdk1 and localizes to the cell cortex and retraction fibers, where it forms complexes with EB1 and p150(glued)/dynactin to stabilize cortical and astral microtubule attachments and orient the spindle; its depletion causes mitotic arrest, reduced inter-kinetochore tension, chromosome misalignment, and randomized spindle orientation [PMID:23509069, PMID:23574715]. MISP acts within a defined cortical pathway: it functions downstream of ezrin—which competes with MISP for cortical actin-binding sites—and upstream of NuMA, and it recruits IQGAP1 to engage active Cdc42 and control p150(glued) distribution [PMID:29669740, PMID:29679050]. Cell-cycle-dependent phosphorylation differentially tunes its biochemistry, with interphase sites enhancing bundling activity and mitotic sites suppressing actin binding [PMID:34023777]. In transporting epithelia MISP preferentially binds ADP-actin near filament pointed ends, confining it to microvillar rootlets where it drives rootlet elongation and recruits fimbrin, a localization enforced by ezrin exclusion from the distal core bundle [PMID:35443169, PMID:38588808, PMID:37205433]. Beyond the cytoskeleton, MISP binds the SARAH domain of MST1/2 to block their homodimerization and autophosphorylation, suppressing Hippo signaling and sustaining YAP activity, which in turn transcriptionally upregulates MISP in a feedback loop [PMID:40019375, PMID:41249743].","teleology":[{"year":2013,"claim":"Established MISP as a mitotic effector by identifying it as a Plk1/Cdk1 substrate that links the actin cortex to spindle positioning, answering how cortical actin couples to astral microtubules.","evidence":"siRNA depletion, Plk1/Cdk1 phosphorylation assays, Co-IP of MISP with p150(glued) and EB1, live-cell spindle imaging and kinetochore tension measurements in cultured cells","pmids":["23509069","23574715"],"confidence":"High","gaps":["Phospho-site identities and how they alter MISP localization were not mapped","Direct vs. indirect nature of the EB1/p150(glued) interactions not resolved structurally"]},{"year":2014,"claim":"Demonstrated that MISP is a direct multivalent actin-binding protein that bundles F-actin, establishing the biochemical basis for its cytoskeletal role independent of mitosis.","evidence":"In vitro F-actin bundling assays with purified protein plus overexpression/knockdown morphological phenotypes in cultured cells","pmids":["24475924"],"confidence":"High","gaps":["Number and location of actin-binding sites not defined","Filament polarity of bundles not addressed"]},{"year":2018,"claim":"Placed MISP within an ordered cortical spindle-orientation pathway, showing it acts downstream of ezrin (via actin-site competition) and engages IQGAP1–Cdc42 to control NuMA and p150(glued).","evidence":"Co-IP (MISP–ezrin, MISP–IQGAP1, IQGAP1–Cdc42), siRNA epistasis, active-Cdc42 pull-down, IQGAP1 rescue, live imaging of NuMA and astral microtubules","pmids":["29669740","29679050"],"confidence":"High","gaps":["How active Cdc42 feeds back onto microtubule capture not mechanistically detailed","Structural basis of the MISP–IQGAP1 interaction unknown"]},{"year":2021,"claim":"Resolved how the cell cycle reprograms MISP function by mapping phospho-sites that separately tune bundling versus binding activities.","evidence":"Site-directed mutagenesis of S394/S395/S400 and S376/S471/S541, in vitro actin-binding/bundling assays, phosphomutant overexpression with morphology readout","pmids":["34023777"],"confidence":"High","gaps":["Kinases for each interphase/mitotic site not all assigned","Quantitative contribution of each site in vivo not established"]},{"year":2022,"claim":"Defined MISP's epithelial role by showing it localizes to and elongates microvillar actin rootlets and recruits fimbrin, with ezrin competition confining it to the basal pointed-end side.","evidence":"Immunofluorescence/live imaging, in vitro bundling and co-sedimentation with purified protein, siRNA/CRISPR KO with rootlet-length phenotype, ezrin loss-of-function","pmids":["35443169"],"confidence":"High","gaps":["What terminates rootlet elongation not defined","Relationship between mitotic cortical role and epithelial microvillar role not unified"]},{"year":2022,"claim":"Provided in vivo physiological context, showing MISP supports intestinal epithelial recovery from inflammation partly via TGF-β1.","evidence":"MISP-knockout mouse DSS colitis model, RT-qPCR for Tgfb1, crypt histology and Ki67 staining","pmids":["36596561"],"confidence":"Medium","gaps":["Mechanism linking MISP to Tgfb1 expression not reconstituted","Single lab; cell-type-specific contribution unresolved"]},{"year":2024,"claim":"Explained the molecular basis of MISP's nucleotide-state-selective actin engagement, showing it binds ADP-actin near pointed ends and assembles polarity-defined bundles, accounting for its rootlet targeting.","evidence":"In vitro TIRF microscopy with purified MISP and differentially nucleotide-loaded actin, cofilin immunostaining of native intestine, solution vs. substrate-attached bundling assays","pmids":["38588808","37205433"],"confidence":"High","gaps":["Structural determinant of ADP-actin preference unknown","How phospho-regulation intersects with nucleotide-state selectivity not tested"]},{"year":2024,"claim":"Uncovered a non-cytoskeletal signaling function: MISP directly inhibits MST1/2 to suppress Hippo signaling and sustain YAP, forming a YAP→MISP positive feedback loop linked to ferroptosis protection.","evidence":"Co-IP (MISP–MST1/2), R390/391A binding-disruption mutant, in vitro autophosphorylation assay, siRNA with ferroptosis readout, in vivo xenograft","pmids":["40019375","41249743"],"confidence":"High","gaps":["Whether actin binding and MST1/2 binding are mutually exclusive not determined","Structural detail of MISP–SARAH domain contact not solved"]},{"year":2024,"claim":"Extended MISP into additional cancer signaling axes (IQGAP1/STAT3/PD-L1, Wnt/β-catenin, JAK2-STAT3, OIP5), implicating it in immune escape and proliferation.","evidence":"Co-IP of MISP–IQGAP1–STAT3 and MISP–OIP5 complexes, β-catenin nuclear localization Westerns, MISP-deficient mouse tumor models, xenografts","pmids":["37633233","38057358","39278512","38474305"],"confidence":"Medium","gaps":["Direct biochemical mechanism for STAT3/β-catenin activation not reconstituted","Whether these axes depend on MISP's actin or MST1/2 activities unclear"]},{"year":null,"claim":"It remains unresolved how MISP's actin-bundling, spindle-orientation, and MST1/2-inhibitory activities are coordinated within a single cell and whether they share or compete for the same regions of the protein.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structure of MISP or its interaction interfaces","No unified model integrating cytoskeletal and Hippo-signaling roles","Tissue-specific dominance of each function undefined"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[2,5,6,10]},{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[9]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[9,3,4]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,4]}],"localization":[{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[1,2,6]},{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[0,3,6]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[12]}],"pathway":[{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[0,1,3,4]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[9,13]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[7,8,11,9]}],"complexes":[],"partners":["EB1","DCTN1","IQGAP1","EZR","MST1","STK4","NUMA1","OIP5"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q8IVT2","full_name":"Mitotic interactor and substrate of PLK1","aliases":["Mitotic spindle positioning protein"],"length_aa":679,"mass_kda":75.4,"function":"Plays a role in mitotic spindle orientation and mitotic progression. Regulates the distribution of dynactin at the cell cortex in a PLK1-dependent manner, thus stabilizing cortical and astral microtubule attachments required for proper mitotic spindle positioning. May link microtubules to the actin cytospkeleton and focal adhesions. May be required for directed cell migration and centrosome orientation. May also be necessary for proper stacking of the Golgi apparatus","subcellular_location":"Cell junction, focal adhesion; Cytoplasm, cytoskeleton; Cytoplasm, cell cortex","url":"https://www.uniprot.org/uniprotkb/Q8IVT2/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/MISP","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/MISP","total_profiled":1310},"omim":[{"mim_id":"615289","title":"MITOTIC SPINDLE-POSITIONING PROTEIN; MISP","url":"https://www.omim.org/entry/615289"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Enhanced","locations":[{"location":"Plasma membrane","reliability":"Enhanced"},{"location":"Focal adhesion sites","reliability":"Enhanced"}],"tissue_specificity":"Tissue enriched","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"intestine","ntpm":185.2}],"url":"https://www.proteinatlas.org/search/MISP"},"hgnc":{"alias_symbol":["DKFZp686H18209","Caprice","MISP1"],"prev_symbol":["C19orf21"]},"alphafold":{"accession":"Q8IVT2","domains":[],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8IVT2","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q8IVT2-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q8IVT2-F1-predicted_aligned_error_v6.png","plddt_mean":53.44},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=MISP","jax_strain_url":"https://www.jax.org/strain/search?query=MISP"},"sequence":{"accession":"Q8IVT2","fasta_url":"https://rest.uniprot.org/uniprotkb/Q8IVT2.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q8IVT2/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8IVT2"}},"corpus_meta":[{"pmid":"23509069","id":"PMC_23509069","title":"MISP is a novel Plk1 substrate required for proper spindle orientation and mitotic progression.","date":"2013","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/23509069","citation_count":63,"is_preprint":false},{"pmid":"23574715","id":"PMC_23574715","title":"The novel actin/focal adhesion-associated protein MISP is involved in mitotic spindle positioning in human cells.","date":"2013","source":"Cell cycle (Georgetown, Tex.)","url":"https://pubmed.ncbi.nlm.nih.gov/23574715","citation_count":41,"is_preprint":false},{"pmid":"37633233","id":"PMC_37633233","title":"Mechanism of M2 type macrophage-derived extracellular vesicles regulating PD-L1 expression via the MISP/IQGAP1 axis in hepatocellular carcinoma immunotherapy resistance.","date":"2023","source":"International immunopharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/37633233","citation_count":31,"is_preprint":false},{"pmid":"28288560","id":"PMC_28288560","title":"Duplication and concerted evolution of MiSp-encoding genes underlie the material properties of minor ampullate silks of cobweb weaving spiders.","date":"2017","source":"BMC evolutionary biology","url":"https://pubmed.ncbi.nlm.nih.gov/28288560","citation_count":31,"is_preprint":false},{"pmid":"35443169","id":"PMC_35443169","title":"Mitotic Spindle Positioning (MISP) is an actin bundler that selectively stabilizes the rootlets of epithelial microvilli.","date":"2022","source":"Cell reports","url":"https://pubmed.ncbi.nlm.nih.gov/35443169","citation_count":22,"is_preprint":false},{"pmid":"29669740","id":"PMC_29669740","title":"Activated ezrin controls MISP levels to ensure correct NuMA polarization and spindle orientation.","date":"2018","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/29669740","citation_count":22,"is_preprint":false},{"pmid":"29679050","id":"PMC_29679050","title":"MISP regulates the IQGAP1/Cdc42 complex to collectively orchestrate spindle orientation and mitotic progression.","date":"2018","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/29679050","citation_count":17,"is_preprint":false},{"pmid":"24475924","id":"PMC_24475924","title":"Caprice/MISP is a novel F-actin bundling protein critical for actin-based cytoskeletal reorganizations.","date":"2014","source":"Genes to cells : devoted to molecular & cellular mechanisms","url":"https://pubmed.ncbi.nlm.nih.gov/24475924","citation_count":15,"is_preprint":false},{"pmid":"40019375","id":"PMC_40019375","title":"MISP Suppresses Ferroptosis via MST1/2 Kinases to Facilitate YAP Activation in Non-Small Cell Lung Cancer.","date":"2025","source":"Advanced science (Weinheim, Baden-Wurttemberg, Germany)","url":"https://pubmed.ncbi.nlm.nih.gov/40019375","citation_count":14,"is_preprint":false},{"pmid":"26706076","id":"PMC_26706076","title":"CAPRICE family genes control flowering time through both promoting and repressing CONSTANS and FLOWERING LOCUS T expression.","date":"2015","source":"Plant science : an international journal of experimental plant biology","url":"https://pubmed.ncbi.nlm.nih.gov/26706076","citation_count":13,"is_preprint":false},{"pmid":"36674734","id":"PMC_36674734","title":"Characteristic Evaluation of Recombinant MiSp/Poly(lactic-co-glycolic) Acid (PLGA) Nanofiber Scaffolds as Potential Scaffolds for Bone Tissue Engineering.","date":"2023","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/36674734","citation_count":8,"is_preprint":false},{"pmid":"38057358","id":"PMC_38057358","title":"PLK1 and its substrate MISP facilitate intrahepatic cholangiocarcinoma progression by promoting lymphatic invasion and impairing E-cadherin adherens junctions.","date":"2023","source":"Cancer gene therapy","url":"https://pubmed.ncbi.nlm.nih.gov/38057358","citation_count":7,"is_preprint":false},{"pmid":"39278512","id":"PMC_39278512","title":"MISP-mediated enhancement of pancreatic cancer growth through the Wnt/β-catenin signaling pathway is suppressed by Fisetin.","date":"2024","source":"Biochimica et biophysica acta. Molecular basis of disease","url":"https://pubmed.ncbi.nlm.nih.gov/39278512","citation_count":7,"is_preprint":false},{"pmid":"38474305","id":"PMC_38474305","title":"Mitotic Spindle Positioning (MISP) Facilitates Colorectal Cancer Progression by Forming a Complex with Opa Interacting Protein 5 (OIP5) and Activating the JAK2-STAT3 Signaling Pathway.","date":"2024","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/38474305","citation_count":6,"is_preprint":false},{"pmid":"36596561","id":"PMC_36596561","title":"Mitotic spindle positioning protein (MISP) deficiency exacerbates dextran sulfate sodium (DSS)-induced colitis in mice.","date":"2022","source":"The Journal of veterinary medical science","url":"https://pubmed.ncbi.nlm.nih.gov/36596561","citation_count":6,"is_preprint":false},{"pmid":"38785491","id":"PMC_38785491","title":"MISP Is Overexpressed in Intestinal Metaplasia and Gastric Cancer.","date":"2024","source":"Current oncology (Toronto, Ont.)","url":"https://pubmed.ncbi.nlm.nih.gov/38785491","citation_count":5,"is_preprint":false},{"pmid":"26039466","id":"PMC_26039466","title":"Overexpressing CAPRICE and GLABRA3 did not change the anthocyanin content of tomato (Solanum lycopersicum) fruit peel.","date":"2015","source":"Plant signaling & behavior","url":"https://pubmed.ncbi.nlm.nih.gov/26039466","citation_count":5,"is_preprint":false},{"pmid":"36996244","id":"PMC_36996244","title":"The Trypanosoma brucei MISP family of invariant proteins is co-expressed with BARP as triple helical bundle structures on the surface of salivary gland forms, but is dispensable for parasite development within the tsetse vector.","date":"2023","source":"PLoS pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/36996244","citation_count":4,"is_preprint":false},{"pmid":"38588808","id":"PMC_38588808","title":"Mitotic spindle positioning protein (MISP) preferentially binds to aged F-actin.","date":"2024","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/38588808","citation_count":1,"is_preprint":false},{"pmid":"34023777","id":"PMC_34023777","title":"Modulation of actin-binding and -bundling activities of MISP/Caprice by multiple phosphorylation.","date":"2021","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/34023777","citation_count":1,"is_preprint":false},{"pmid":"40436766","id":"PMC_40436766","title":"The predictive value of MISP for the postoperative outcomes of gastric cancer.","date":"2025","source":"Biomedical research (Tokyo, Japan)","url":"https://pubmed.ncbi.nlm.nih.gov/40436766","citation_count":1,"is_preprint":false},{"pmid":"37205433","id":"PMC_37205433","title":"Mitotic spindle positioning protein (MISP) is an actin bundler that senses ADP-actin and binds near the pointed ends of filaments.","date":"2023","source":"bioRxiv : the preprint server for biology","url":"https://pubmed.ncbi.nlm.nih.gov/37205433","citation_count":0,"is_preprint":false},{"pmid":"41249743","id":"PMC_41249743","title":"MISP promotes the progression of lung adenocarcinoma through Inhibition of the Hippo signaling pathway.","date":"2025","source":"Discover oncology","url":"https://pubmed.ncbi.nlm.nih.gov/41249743","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.11.03.25339450","title":"Patient-Level Stroke Outcomes Following Linkage of a Statewide Stroke Registry to Administrative Claims Data","date":"2025-11-06","source":"bioRxiv","url":"https://doi.org/10.1101/2025.11.03.25339450","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":13467,"output_tokens":4635,"usd":0.054963,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":12582,"output_tokens":3641,"usd":0.076967,"stage2_stop_reason":"end_turn"},"total_usd":0.13193,"stage1_batch_id":"msgbatch_01BbTCMyrRPyhtEq6b1MdwCN","stage2_batch_id":"msgbatch_011dEed5bbxYvTAaQWKJevxu","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2013,\n      \"finding\": \"MISP (C19orf21) is a substrate of Plk1; Plk1 phosphorylates MISP, and this phosphorylation is required for the metaphase-to-anaphase transition and proper mitotic spindle positioning. MISP is an actin-associated protein that forms a complex with p150(glued) (a dynein-dynactin subunit) and regulates its cortical distribution, thereby stabilizing cortical and astral microtubule attachments.\",\n      \"method\": \"Loss-of-function (siRNA depletion), phosphorylation assays identifying Plk1 as the kinase, co-immunoprecipitation of MISP with p150(glued), live-cell imaging of spindle orientation and astral microtubule length\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, kinase substrate assay, KD with defined spindle phenotype; replicated independently in a second concurrent study (PMID:23574715)\",\n      \"pmids\": [\"23509069\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"MISP associates with the actin cytoskeleton and focal adhesions and is expressed only in adherent cell types. During mitosis, MISP is phosphorylated by Cdk1 and localizes to retraction fibers. MISP interacts with the +TIP protein EB1 and with p150(glued) (dynein/dynactin). Depletion of MISP causes mitotic arrest with reduced tension across sister kinetochores, chromosome misalignment, spindle multipolarity (in cells with supernumerary centrosomes), and randomized spindle orientation.\",\n      \"method\": \"Co-immunoprecipitation (MISP with EB1 and p150glued), Cdk1 phosphorylation assay, siRNA depletion with live-cell imaging, immunofluorescence of focal adhesions and retraction fibers, kinetochore tension measurements\",\n      \"journal\": \"Cell cycle (Georgetown, Tex.)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, kinase assay, KD with multiple orthogonal phenotypic readouts; independent replication of core findings (PMID:23509069)\",\n      \"pmids\": [\"23574715\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"MISP/Caprice is a direct actin-binding protein with multiple actin-binding sites that forms mesh-like F-actin bundles in vitro. Overexpression induces stress fiber-like thick filaments and knockdown causes filopodial formation, demonstrating a direct role in actin cytoskeletal organization. It is expressed in cells/tissues with specialized actin structures such as growth cones and inner ear hair cell stereocilia.\",\n      \"method\": \"In vitro F-actin bundling assay with purified protein, overexpression and knockdown in cultured cells with morphological readout, fractionation identifying MISP in highly insoluble scaffold fraction\",\n      \"journal\": \"Genes to cells : devoted to molecular & cellular mechanisms\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro bundling assay with purified protein plus loss- and gain-of-function cellular phenotypes in a single study\",\n      \"pmids\": [\"24475924\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"MISP functions downstream of the ERM family member ezrin and upstream of NuMA in the spindle orientation pathway. SLK/LOK-activated (phosphorylated) ezrin competes with MISP for actin-binding sites at the cell cortex, limiting cortical MISP levels. Excessive MISP accumulation disrupts crescent-like NuMA polarization at the cortex and reduces astral microtubule dynamics. MISP directly interacts with ezrin.\",\n      \"method\": \"Co-immunoprecipitation (MISP–ezrin interaction), siRNA epistasis experiments (SLK/LOK, ezrin, MISP), live-cell imaging of NuMA localization and astral microtubule dynamics, cortical fractionation\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct Co-IP, genetic epistasis with defined pathway ordering, multiple orthogonal phenotypic readouts in a single study\",\n      \"pmids\": [\"29669740\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"MISP interacts with the scaffolding protein IQGAP1, and through IQGAP1 binds the active (GTP-bound) form of Cdc42. MISP depletion increases IQGAP1 cortical accumulation and decreases Cdc42 activity. IQGAP1 overexpression rescues MISP-depletion phenotypes (spindle misorientation, loss of astral microtubules, prolonged mitosis) and restores active Cdc42 levels, placing IQGAP1 downstream of MISP. IQGAP1 also acts downstream of MISP in controlling p150(glued) localization.\",\n      \"method\": \"Co-immunoprecipitation (MISP–IQGAP1, IQGAP1–Cdc42), siRNA depletion epistasis, active-Cdc42 pull-down assay, IQGAP1 overexpression rescue experiments, immunofluorescence of p150(glued) localization\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal Co-IP, active-Cdc42 pull-down, rescue epistasis, multiple orthogonal phenotypic readouts in one study\",\n      \"pmids\": [\"29679050\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Phosphorylation of MISP at S394, S395, and S400 increases actin-bundling activity (but not actin-binding activity), inducing stress fiber formation in interphase cells. Mitotic phosphorylation at S376, S471, and S541 suppresses actin-binding activity. Thus, cell-cycle-dependent phosphorylation differentially regulates MISP actin-binding versus bundling activities.\",\n      \"method\": \"Site-directed mutagenesis of phosphorylation sites, in vitro actin-binding and bundling assays, overexpression of phosphomutants in cells with morphological readout\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro biochemical assays combined with mutagenesis and cellular phenotype in a single study\",\n      \"pmids\": [\"34023777\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"MISP localizes specifically to the rootlet end (basal, pointed-end side) of microvillar actin core bundles in transporting epithelia. Purified MISP exhibits potent F-actin bundling activity in vitro and promotes rootlet elongation in cells. MISP-bundled filaments recruit fimbrin, which further elongates and stabilizes bundles. Ezrin prevents MISP from decorating the membrane-wrapped distal end of the core bundle, confining MISP to the rootlet.\",\n      \"method\": \"Immunofluorescence/live imaging for localization, in vitro actin bundling assay with purified protein, siRNA/CRISPR KO with microvillar phenotype readout (rootlet length), co-sedimentation assays, ezrin loss-of-function experiments\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro reconstitution with purified protein, cellular loss-of-function with defined morphological phenotype, mechanistic dissection of ezrin competition; multiple orthogonal methods\",\n      \"pmids\": [\"35443169\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"MISP downregulation by M2 macrophage-derived extracellular vesicles promotes IQGAP1 nuclear translocation and activates STAT3 phosphorylation in hepatocellular carcinoma cells, leading to upregulation of PD-L1 and immune escape. MISP, IQGAP1, and STAT3 interact in a complex as shown by co-immunoprecipitation. MISP negatively correlates with IQGAP1/PD-L1 expression.\",\n      \"method\": \"Co-immunoprecipitation (MISP–IQGAP1–STAT3 complex), MISP/IQGAP1 overexpression experiments, flow cytometry of CD8+ T cells, ELISA for cytokines, in vivo nude mouse xenograft\",\n      \"journal\": \"International immunopharmacology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — Co-IP complex validated, in vivo rescue experiment, but single lab with limited mechanistic depth on MISP's direct biochemical role\",\n      \"pmids\": [\"37633233\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"In intrahepatic cholangiocarcinoma cells, MISP localizes to adherens junctions and suppresses E-cadherin dimerization. PLK1 phosphorylates MISP (confirmed as a PLK1 substrate in this context) and, together with MISP, disrupts adherens junctions to promote lymphatic invasion and cell motility.\",\n      \"method\": \"siRNA knockdown of PLK1 or MISP with trans-lymphatic endothelial migration and wound healing assays, immunofluorescence of focal adhesions and adherens junctions, in vivo tumorigenesis assay\",\n      \"journal\": \"Cancer gene therapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — KD with defined cellular phenotypes, localization by IF, single lab without in vitro reconstitution of E-cadherin dimerization suppression\",\n      \"pmids\": [\"38057358\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"MISP directly binds the SARAH domain of MST1/2 kinases, inhibiting their homodimerization and autophosphorylation, thereby sustaining YAP activation (Hippo pathway inhibition). YAP activation in turn increases SLC7A11 expression to protect cells from ferroptosis. A MISP-R390/391A mutant that disrupts MISP–MST1/2 binding abolishes this effect. YAP also transcriptionally activates MISP, creating a positive feedback loop.\",\n      \"method\": \"Co-immunoprecipitation (MISP–MST1/2), site-directed mutagenesis (R390/391A), in vitro kinase/autophosphorylation assay, siRNA knockdown with ferroptosis sensitivity readout, Western blot for Hippo pathway components, in vivo xenograft\",\n      \"journal\": \"Advanced science (Weinheim, Baden-Wurttemberg, Germany)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — Co-IP, mutagenesis disrupting binding, in vitro kinase assay, multiple orthogonal readouts in one study with in vivo validation\",\n      \"pmids\": [\"40019375\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"MISP preferentially binds ADP-actin (aged/pointed-end-enriched) filaments over ADP-Pi-actin filaments, and in vitro TIRF assays show MISP binds at or near the pointed ends of growing actin filaments. In solution, MISP assembles parallel bundles of uniform polarity, whereas substrate-attached MISP can bundle filaments in both parallel and antiparallel configurations. Microvillar rootlets are decorated with cofilin (indicating high ADP-actin content), consistent with MISP's nucleotide-state preference driving its rootlet localization.\",\n      \"method\": \"In vitro TIRF microscopy with purified MISP and differentially nucleotide-loaded actin filaments, immunostaining of native intestinal tissue for cofilin, in vitro bundling assays in solution vs. substrate-attached conditions\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro reconstitution with purified protein under multiple controlled conditions, corroborated by native tissue immunostaining; also supported by companion preprint (PMID:37205433)\",\n      \"pmids\": [\"38588808\", \"37205433\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"MISP promotes nuclear accumulation of β-catenin, thereby activating the Wnt/β-catenin signaling pathway in pancreatic cancer cells. Fisetin suppresses MISP expression by downregulating the transcription factor MYB, identifying MYB as a transcriptional activator of MISP.\",\n      \"method\": \"Overexpression/knockdown of MISP with Western blot for β-catenin nuclear localization, in vitro proliferation assays, in vivo xenograft, drug screening library\",\n      \"journal\": \"Biochimica et biophysica acta. Molecular basis of disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Weak — cellular overexpression/KD with β-catenin localization readout, single lab without direct biochemical reconstitution of the MISP–β-catenin interaction\",\n      \"pmids\": [\"39278512\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"MISP forms a complex with Opa Interacting Protein 5 (OIP5) in the cytoplasm, and MISP influences OIP5 expression in a unidirectional manner. MISP also increases phosphorylated STAT3 levels via the JAK2-STAT3 signaling pathway in colorectal cancer cells.\",\n      \"method\": \"Co-immunoprecipitation (MISP–OIP5 complex), Western blot for pSTAT3, MISP-deficient mouse model (AOM/DSS colitis-induced tumors), siRNA knockdown with cell proliferation readout\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — Co-IP for complex, in vivo KO model with tumor phenotype, but limited mechanistic depth on how MISP activates JAK2-STAT3; single lab\",\n      \"pmids\": [\"38474305\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"MISP directly interacts with MST1 (co-immunoprecipitation confirmed), inhibiting Hippo signaling and leading to increased YAP, TAZ, and CYR61 expression and decreased MST1 and phospho-YAP levels in lung adenocarcinoma cells.\",\n      \"method\": \"Co-immunoprecipitation (MISP–MST1), Western blot for Hippo pathway components, overexpression/knockdown with in vitro and in vivo proliferation and migration assays\",\n      \"journal\": \"Discover oncology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — Co-IP confirming direct interaction with MST1, cellular phenotype with pathway readout, but single lab and limited mechanistic depth; largely corroborates PMID:40019375\",\n      \"pmids\": [\"41249743\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"MISP-deficient mice show exacerbated DSS-induced colitis, with decreased intestinal epithelial cell proliferation and significantly reduced Tgfb1 (TGF-β1) anti-inflammatory cytokine expression in the colon, indicating MISP supports colon recovery from inflammation partly through TGF-β1 signaling.\",\n      \"method\": \"MISP-knockout mouse model (DSS colitis), RT-qPCR for Tgfb1 and other cytokines, histological assessment of crypt morphology, Ki67 proliferation staining\",\n      \"journal\": \"The Journal of veterinary medical science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — defined KO model with specific molecular readout (Tgfb1), single lab without mechanistic reconstitution of how MISP regulates TGF-β1\",\n      \"pmids\": [\"36596561\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"MISP (C19orf21/Caprice) is a cell-cycle-regulated actin-binding and F-actin bundling protein that, during mitosis, is phosphorylated by Plk1 and Cdk1 to localize to the cell cortex and retraction fibers where it interacts with EB1, p150(glued)/dynactin, and IQGAP1–Cdc42 to stabilize astral microtubule attachments and orient the mitotic spindle; in interphase epithelial cells, MISP preferentially binds ADP-actin at filament pointed ends to bundle rootlet actin in microvilli, a localization enforced by competition with ezrin at the distal core bundle; beyond cytoskeletal roles, MISP inhibits MST1/2 homodimerization and autophosphorylation to suppress Hippo signaling and sustain YAP activation (which in turn transcriptionally upregulates MISP), and MISP additionally modulates IQGAP1/STAT3/PD-L1 and Wnt/β-catenin pathways in cancer contexts.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"MISP (C19orf21/Caprice) is a cell-cycle-regulated, direct actin-binding and F-actin-bundling protein that couples cortical actin organization to mitotic spindle positioning and, in epithelia, to microvillar architecture [#0, #2, #6]. During mitosis it is phosphorylated by Plk1 and Cdk1 and localizes to the cell cortex and retraction fibers, where it forms complexes with EB1 and p150(glued)/dynactin to stabilize cortical and astral microtubule attachments and orient the spindle; its depletion causes mitotic arrest, reduced inter-kinetochore tension, chromosome misalignment, and randomized spindle orientation [#0, #1]. MISP acts within a defined cortical pathway: it functions downstream of ezrin\\u2014which competes with MISP for cortical actin-binding sites\\u2014and upstream of NuMA, and it recruits IQGAP1 to engage active Cdc42 and control p150(glued) distribution [#3, #4]. Cell-cycle-dependent phosphorylation differentially tunes its biochemistry, with interphase sites enhancing bundling activity and mitotic sites suppressing actin binding [#5]. In transporting epithelia MISP preferentially binds ADP-actin near filament pointed ends, confining it to microvillar rootlets where it drives rootlet elongation and recruits fimbrin, a localization enforced by ezrin exclusion from the distal core bundle [#6, #10]. Beyond the cytoskeleton, MISP binds the SARAH domain of MST1/2 to block their homodimerization and autophosphorylation, suppressing Hippo signaling and sustaining YAP activity, which in turn transcriptionally upregulates MISP in a feedback loop [#9, #13].\",\n  \"teleology\": [\n    {\n      \"year\": 2013,\n      \"claim\": \"Established MISP as a mitotic effector by identifying it as a Plk1/Cdk1 substrate that links the actin cortex to spindle positioning, answering how cortical actin couples to astral microtubules.\",\n      \"evidence\": \"siRNA depletion, Plk1/Cdk1 phosphorylation assays, Co-IP of MISP with p150(glued) and EB1, live-cell spindle imaging and kinetochore tension measurements in cultured cells\",\n      \"pmids\": [\"23509069\", \"23574715\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Phospho-site identities and how they alter MISP localization were not mapped\", \"Direct vs. indirect nature of the EB1/p150(glued) interactions not resolved structurally\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Demonstrated that MISP is a direct multivalent actin-binding protein that bundles F-actin, establishing the biochemical basis for its cytoskeletal role independent of mitosis.\",\n      \"evidence\": \"In vitro F-actin bundling assays with purified protein plus overexpression/knockdown morphological phenotypes in cultured cells\",\n      \"pmids\": [\"24475924\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Number and location of actin-binding sites not defined\", \"Filament polarity of bundles not addressed\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Placed MISP within an ordered cortical spindle-orientation pathway, showing it acts downstream of ezrin (via actin-site competition) and engages IQGAP1\\u2013Cdc42 to control NuMA and p150(glued).\",\n      \"evidence\": \"Co-IP (MISP\\u2013ezrin, MISP\\u2013IQGAP1, IQGAP1\\u2013Cdc42), siRNA epistasis, active-Cdc42 pull-down, IQGAP1 rescue, live imaging of NuMA and astral microtubules\",\n      \"pmids\": [\"29669740\", \"29679050\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How active Cdc42 feeds back onto microtubule capture not mechanistically detailed\", \"Structural basis of the MISP\\u2013IQGAP1 interaction unknown\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Resolved how the cell cycle reprograms MISP function by mapping phospho-sites that separately tune bundling versus binding activities.\",\n      \"evidence\": \"Site-directed mutagenesis of S394/S395/S400 and S376/S471/S541, in vitro actin-binding/bundling assays, phosphomutant overexpression with morphology readout\",\n      \"pmids\": [\"34023777\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Kinases for each interphase/mitotic site not all assigned\", \"Quantitative contribution of each site in vivo not established\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Defined MISP's epithelial role by showing it localizes to and elongates microvillar actin rootlets and recruits fimbrin, with ezrin competition confining it to the basal pointed-end side.\",\n      \"evidence\": \"Immunofluorescence/live imaging, in vitro bundling and co-sedimentation with purified protein, siRNA/CRISPR KO with rootlet-length phenotype, ezrin loss-of-function\",\n      \"pmids\": [\"35443169\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"What terminates rootlet elongation not defined\", \"Relationship between mitotic cortical role and epithelial microvillar role not unified\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Provided in vivo physiological context, showing MISP supports intestinal epithelial recovery from inflammation partly via TGF-\\u03b21.\",\n      \"evidence\": \"MISP-knockout mouse DSS colitis model, RT-qPCR for Tgfb1, crypt histology and Ki67 staining\",\n      \"pmids\": [\"36596561\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism linking MISP to Tgfb1 expression not reconstituted\", \"Single lab; cell-type-specific contribution unresolved\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Explained the molecular basis of MISP's nucleotide-state-selective actin engagement, showing it binds ADP-actin near pointed ends and assembles polarity-defined bundles, accounting for its rootlet targeting.\",\n      \"evidence\": \"In vitro TIRF microscopy with purified MISP and differentially nucleotide-loaded actin, cofilin immunostaining of native intestine, solution vs. substrate-attached bundling assays\",\n      \"pmids\": [\"38588808\", \"37205433\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural determinant of ADP-actin preference unknown\", \"How phospho-regulation intersects with nucleotide-state selectivity not tested\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Uncovered a non-cytoskeletal signaling function: MISP directly inhibits MST1/2 to suppress Hippo signaling and sustain YAP, forming a YAP\\u2192MISP positive feedback loop linked to ferroptosis protection.\",\n      \"evidence\": \"Co-IP (MISP\\u2013MST1/2), R390/391A binding-disruption mutant, in vitro autophosphorylation assay, siRNA with ferroptosis readout, in vivo xenograft\",\n      \"pmids\": [\"40019375\", \"41249743\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether actin binding and MST1/2 binding are mutually exclusive not determined\", \"Structural detail of MISP\\u2013SARAH domain contact not solved\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Extended MISP into additional cancer signaling axes (IQGAP1/STAT3/PD-L1, Wnt/\\u03b2-catenin, JAK2-STAT3, OIP5), implicating it in immune escape and proliferation.\",\n      \"evidence\": \"Co-IP of MISP\\u2013IQGAP1\\u2013STAT3 and MISP\\u2013OIP5 complexes, \\u03b2-catenin nuclear localization Westerns, MISP-deficient mouse tumor models, xenografts\",\n      \"pmids\": [\"37633233\", \"38057358\", \"39278512\", \"38474305\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct biochemical mechanism for STAT3/\\u03b2-catenin activation not reconstituted\", \"Whether these axes depend on MISP's actin or MST1/2 activities unclear\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"It remains unresolved how MISP's actin-bundling, spindle-orientation, and MST1/2-inhibitory activities are coordinated within a single cell and whether they share or compete for the same regions of the protein.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structure of MISP or its interaction interfaces\", \"No unified model integrating cytoskeletal and Hippo-signaling roles\", \"Tissue-specific dominance of each function undefined\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [2, 5, 6, 10]},\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [9]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [9, 3, 4]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 4]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [1, 2, 6]},\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [0, 3, 6]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [12]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [0, 1, 3, 4]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [9, 13]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [7, 8, 11, 9]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"EB1\", \"DCTN1\", \"IQGAP1\", \"EZR\", \"MST1\", \"STK4\", \"NUMA1\", \"OIP5\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}