{"gene":"MICAL3","run_date":"2026-06-10T02:59:50","timeline":{"discoveries":[{"year":2011,"finding":"MICAL3 interacts with both Rab8A and ELKS, linking these two proteins. Rab8A associates with exocytotic vesicles in a Rab6-dependent manner, and MICAL3 acts as a scaffold connecting Rab8A and ELKS at the cell cortex. Expression of a MICAL3 mutant with an inactive monooxygenase domain caused strong accumulation of secretory vesicles docked at the cell cortex that failed to fuse with the plasma membrane, indicating that MICAL3's monooxygenase activity is required for vesicle-docking complex remodeling and fusion.","method":"Co-immunoprecipitation, live-cell imaging of vesicle dynamics, dominant-negative monooxygenase mutant expression, FRAP","journal":"Current biology : CB","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, dominant-negative mutant with specific phenotypic readout, live imaging; replicated by subsequent studies","pmids":["21596566"],"is_preprint":false},{"year":2005,"finding":"MICAL3 interacts with Rab1 in yeast two-hybrid and GST pulldown experiments. MICAL3 localizes to a microtubule-associated compartment, as treatment with the microtubule-depolymerizing drug nocodazole disrupts this localization, indicating a link between MICAL3 and the microtubule cytoskeleton.","method":"Yeast two-hybrid, GST pulldown, immunofluorescence with nocodazole treatment","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — yeast two-hybrid and GST pulldown for interaction, single lab, drug perturbation for localization","pmids":["15694364"],"is_preprint":false},{"year":2007,"finding":"Knockdown of MICAL3 in chick motor neurons via RNAi caused ectopic positioning of motor neuron cell bodies in the peripheral nervous system, placing MICAL3 downstream of Neuropilin-2/Plexin-A2 semaphorin signaling as a cytoplasmic effector that controls somal positioning through cytoskeletal reorganization.","method":"RNA interference in chick embryo, analysis of motor neuron positioning","journal":"Neural development","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss-of-function RNAi with specific cellular phenotype, epistasis placement downstream of semaphorin/plexin, single lab","pmids":["17971221"],"is_preprint":false},{"year":2012,"finding":"MICAL3 (as MICAL-3) was shown to interact with Rab1 and Rab35 effector binding is only slightly inhibited by phosphocholination, in contrast to GEF and GDI interactions which are strongly inhibited, demonstrating that MICAL-3 functions as a Rab effector protein.","method":"Biochemical binding assays with phosphocholinated Rab proteins, quantitative interaction measurements","journal":"The EMBO journal","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vitro biochemical assay with modified Rab proteins, single lab, defines MICAL3 as Rab effector","pmids":["22307087"],"is_preprint":false},{"year":2012,"finding":"Human MICAL3, like MICAL1 and MICAL2, is required for normal actin stress fiber organization in non-neural cells, and this actin-regulatory function depends on its generation of reactive oxygen species (ROS) via the monooxygenase domain.","method":"siRNA knockdown, ROS inhibitor treatment, immunofluorescence of actin stress fibers","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss-of-function with specific cytoskeletal phenotype and ROS dependency demonstrated, single lab","pmids":["22331357"],"is_preprint":false},{"year":2015,"finding":"MICAL3 was identified as part of a NINL-associated interactome at the base of cilia, and interacts with Rab8 to regulate vesicle docking and fusion for cilia-directed cargo delivery. Genetic interaction between NINL and CC2D2A was demonstrated in zebrafish, and MICAL3 was linked to this pathway.","method":"Co-immunoprecipitation/interactome analysis, zebrafish morpholino knockdown, immunolocalization","journal":"PLoS genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP interactome, in vivo genetic interaction in zebrafish, localization data; single lab","pmids":["26485645"],"is_preprint":false},{"year":2016,"finding":"MICAL3 forms a direct complex with the centralspindlin component MKLP1, is recruited to the central spindle and midbody during cytokinesis, and targets ELKS and Rab8A-positive vesicles to the midbody. Knockout of MICAL3 increases cytokinetic failure frequency and delays abscission. This scaffolding function is independent of MICAL3's enzymatic monooxygenase activity.","method":"Cross-linking mass spectrometry, MICAL3 knockout, cell biological assays (cytokinesis failure quantification, abscission timing), immunofluorescence","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — cross-linking MS for direct interaction, knockout with defined cellular phenotype, mechanistic dissection of enzymatic vs. scaffolding function; multiple orthogonal methods in one study","pmids":["27528609"],"is_preprint":false},{"year":2017,"finding":"Cross-linking mass spectrometry combined with deletion analysis defined the minimal binding domains of the MICAL3/ELKS/Rab8A complex involved in exocytosis, providing structural constraints on this protein assembly.","method":"Cross-linking mass spectrometry (XL-MS), deletion analysis, biochemical interaction assays","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 1 / Moderate — XL-MS with deletion validation defines binding interfaces; single lab, proof-of-concept study","pmids":["29044157"],"is_preprint":false},{"year":2018,"finding":"Human MICAL3 directly associates with F-actin, which activates its catalytic activity. MICAL3 uses NADPH-dependent redox activity to oxidize actin methionine residues M44/M47, dismantling filaments and limiting polymerization. MsrB/SelR reductase enzymes counteract MICAL3's effect on F-actin both in vitro and in vivo.","method":"In vitro F-actin binding and disassembly assays, NADPH consumption assays, genetic experiments in Drosophila, mass spectrometry of oxidized actin residues","journal":"Scientific reports","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro reconstitution with purified proteins, site-specific modification identified by MS, genetic validation in vivo, reversibility demonstrated; multiple orthogonal methods","pmids":["29343822"],"is_preprint":false},{"year":2018,"finding":"Semaphorin 3 stimulation induces interaction among MICAL3, CRMP2, and Numb in breast cancer stem-like cells. MICAL3 monooxygenase activity (MO activity) is required for this interaction, for Numb protein accumulation, and for symmetric cell division. MICAL3 knockdown decreases sphere formation and shifts CSCs from symmetric to asymmetric division.","method":"Co-immunoprecipitation, MICAL3 siRNA knockdown, sphere formation assay, division mode quantification (symmetric vs. asymmetric)","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, loss-of-function with quantitative division mode phenotype, enzymatic activity requirement tested; single lab","pmids":["30587593"],"is_preprint":false},{"year":2020,"finding":"Crystal structure of human MICAL3 FMO and calponin-homology (CH) domains was solved. MICAL3 contains an FAD/NADP-binding Rossmann-fold domain for monooxygenase activity. Kinetic analysis showed that F-actin dramatically increases MICAL3 catalytic efficiency only when the CH domain is present. Mutation of Glu213 in the FMO domain and Arg530 in the CH domain abolished this F-actin-stimulated catalytic activation.","method":"X-ray crystallography, kinetic enzyme assays, site-directed mutagenesis (E213A, R530A)","journal":"IUCrJ","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structure plus kinetic assays plus mutagenesis in single study with multiple orthogonal methods; definitively maps F-actin-dependent activation mechanism","pmids":["31949908"],"is_preprint":false},{"year":2024,"finding":"CHK1 physically interacts with MICAL3 in mouse zygotes, as identified by co-immunoprecipitation and mass spectrometry. Gain-of-function CHK1 mutants enhance interaction with MICAL3 and increase MICAL3 enzymatic activity, causing excessive F-actin depolymerization that disrupts pronuclear envelope breakdown.","method":"Co-immunoprecipitation and mass spectrometry of ~6000 mouse zygotes, gain-of-function CHK1 mutant analysis, F-actin imaging","journal":"EMBO reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP/MS interaction, gain-of-function mutant with specific cellular phenotype; single lab","pmids":["39358552"],"is_preprint":false}],"current_model":"MICAL3 is a flavoprotein monooxygenase that uses NADPH-dependent redox activity to oxidize actin methionine residues M44/M47, directly dismantling F-actin filaments in a process stimulated by F-actin binding to its calponin-homology domain (with Glu213 and Arg530 being critical for this activation); it also functions as a scaffold at the midbody and exocytotic sites by directly binding centralspindlin component MKLP1 and linking Rab8A-positive vesicles to ELKS, thereby coordinating vesicle docking/fusion during exocytosis and cytokinetic abscission, while additionally acting downstream of semaphorin/plexin signaling to regulate cytoskeletal reorganization, symmetric cell division, and pronuclear envelope breakdown through interactions with CRMP2/Numb and CHK1 respectively."},"narrative":{"mechanistic_narrative":"MICAL3 is a flavoprotein monooxygenase that couples NADPH-dependent redox chemistry to direct remodeling of the actin cytoskeleton and to scaffolding of membrane-trafficking machinery [PMID:29343822, PMID:21596566]. As an enzyme, it binds F-actin through its calponin-homology (CH) domain, which dramatically stimulates its catalytic efficiency and allows it to oxidize actin methionine residues M44/M47, dismantling filaments and limiting polymerization; this oxidation is reversed by MsrB/SelR reductases [PMID:29343822]. A crystal structure of the FMO and CH domains shows an FAD/NADP-binding Rossmann fold, and mutation of Glu213 (FMO) or Arg530 (CH) abolishes the F-actin-stimulated activation, defining the structural basis of substrate-coupled catalysis [PMID:31949908]. Independently of its enzymatic activity, MICAL3 acts as a Rab effector and trafficking scaffold: it links Rab8A-positive exocytotic vesicles to ELKS at the cell cortex, where its monooxygenase activity is required for remodeling the docking complex to permit vesicle fusion [PMID:21596566, PMID:22307087]. The same scaffolding logic operates during cytokinesis, where MICAL3 binds the centralspindlin component MKLP1, is recruited to the central spindle and midbody, and targets ELKS and Rab8A vesicles there to support abscission [PMID:27528609]. Through its actin-regulatory and trafficking functions MICAL3 contributes to broader cellular processes including semaphorin/plexin-directed cytoskeletal reorganization and motor neuron somal positioning [PMID:17971221], symmetric division of cancer stem-like cells via a Semaphorin 3-induced MICAL3/CRMP2/Numb complex [PMID:30587593], cilia-directed cargo delivery [PMID:26485645], and pronuclear envelope breakdown downstream of CHK1 [PMID:39358552].","teleology":[{"year":2005,"claim":"Established the first molecular handle on MICAL3 by placing it at the microtubule cytoskeleton as a candidate Rab1-binding protein, raising the question of whether it functions in membrane trafficking.","evidence":"Yeast two-hybrid and GST pulldown for Rab1 binding, immunofluorescence with nocodazole","pmids":["15694364"],"confidence":"Medium","gaps":["Rab1 interaction not validated by reciprocal endogenous Co-IP","no functional consequence of the microtubule association defined"]},{"year":2007,"claim":"Showed MICAL3 acts as a cytoplasmic effector of semaphorin signaling, answering whether it transduces guidance cues into cytoskeletal control in vivo.","evidence":"RNAi knockdown in chick embryo motor neurons, somal positioning analysis","pmids":["17971221"],"confidence":"Medium","gaps":["molecular link between Neuropilin-2/Plexin-A2 and MICAL3 not biochemically defined","single in vivo system"]},{"year":2011,"claim":"Defined MICAL3 as a scaffold bridging Rab8A vesicles and ELKS at the cortex and showed its monooxygenase activity is required for vesicle fusion, establishing a catalytic role in exocytosis.","evidence":"Reciprocal Co-IP, live-cell vesicle imaging, dominant-negative monooxygenase mutant, FRAP","pmids":["21596566"],"confidence":"High","gaps":["redox substrate at the docking site not identified","structural geometry of the Rab8A-MICAL3-ELKS bridge unresolved"]},{"year":2012,"claim":"Distinguished MICAL3 as a bona fide Rab effector and tied its monooxygenase-dependent ROS production to actin stress fiber organization in non-neural cells.","evidence":"Binding assays with phosphocholinated Rabs; siRNA knockdown with ROS inhibitor and actin immunofluorescence","pmids":["22307087","22331357"],"confidence":"Medium","gaps":["effector-versus-GEF/GDI selectivity tested only biochemically","ROS-dependence inferred from inhibitors rather than direct substrate identification"]},{"year":2015,"claim":"Extended MICAL3's Rab8-coupled trafficking role to the ciliary base, linking it to NINL-directed cargo delivery.","evidence":"Co-IP interactome, zebrafish morpholino knockdown, immunolocalization","pmids":["26485645"],"confidence":"Medium","gaps":["direct versus indirect association with NINL not separated","enzymatic requirement at cilia not tested"]},{"year":2016,"claim":"Resolved that MICAL3's midbody function is structural by showing direct MKLP1 binding and vesicle targeting that supports abscission independently of monooxygenase activity, dissociating scaffolding from catalysis.","evidence":"Cross-linking mass spectrometry, MICAL3 knockout, cytokinesis-failure and abscission-timing assays","pmids":["27528609"],"confidence":"High","gaps":["how MKLP1 recruitment is temporally regulated unknown","whether catalytic activity contributes elsewhere in cytokinesis not addressed"]},{"year":2017,"claim":"Defined the minimal binding interfaces of the MICAL3/ELKS/Rab8A exocytic complex, providing structural constraints on the assembly.","evidence":"Cross-linking mass spectrometry with deletion analysis","pmids":["29044157"],"confidence":"Medium","gaps":["interfaces mapped by XL-MS, not high-resolution structure","stoichiometry of the assembly not determined"]},{"year":2018,"claim":"Established the core enzymatic mechanism: MICAL3 directly binds and oxidizes F-actin at methionine M44/M47 to disassemble filaments, a modification reversed by MsrB/SelR reductases, defining a redox switch on actin.","evidence":"In vitro F-actin binding/disassembly and NADPH assays, MS of oxidized residues, Drosophila genetics","pmids":["29343822"],"confidence":"High","gaps":["regulation of catalysis in the cellular context not fully defined","relationship between actin oxidation and vesicle-docking remodeling unresolved"]},{"year":2018,"claim":"Connected MICAL3 monooxygenase activity to cell-fate control by showing a Semaphorin 3-induced MICAL3/CRMP2/Numb complex drives symmetric division of cancer stem-like cells.","evidence":"Co-IP, siRNA knockdown, sphere formation and symmetric/asymmetric division quantification","pmids":["30587593"],"confidence":"Medium","gaps":["mechanism linking Numb stabilization to MICAL3 catalysis unclear","directness of CRMP2/Numb interactions not established"]},{"year":2020,"claim":"Provided the structural and kinetic basis for substrate-coupled activation, showing F-actin stimulates catalysis only when the CH domain is present and identifying Glu213 and Arg530 as essential for this allostery.","evidence":"X-ray crystallography of FMO/CH domains, kinetic assays, E213A/R530A mutagenesis","pmids":["31949908"],"confidence":"High","gaps":["full-length enzyme structure including regulatory regions not solved","conformational coupling between CH and FMO domains not visualized"]},{"year":2024,"claim":"Identified CHK1 as an upstream regulator of MICAL3 activity in zygotes, where excessive MICAL3-driven actin depolymerization disrupts pronuclear envelope breakdown.","evidence":"Co-IP/MS from mouse zygotes, gain-of-function CHK1 mutants, F-actin imaging","pmids":["39358552"],"confidence":"Medium","gaps":["whether CHK1 directly phosphorylates MICAL3 not shown","single in vivo system, no reconstitution of the regulatory step"]},{"year":null,"claim":"How MICAL3's two activities — actin-oxidizing catalysis and Rab/centralspindlin scaffolding — are spatially and temporally integrated within a single cell remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["no unified model coupling redox actin remodeling to vesicle docking at exocytic and midbody sites","upstream signals selecting between catalytic and scaffolding modes not defined"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0016491","term_label":"oxidoreductase activity","supporting_discovery_ids":[4,8,10]},{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[8]},{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[8,10]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,6]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[0]},{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[1,8]},{"term_id":"GO:0005815","term_label":"microtubule organizing center","supporting_discovery_ids":[6]}],"pathway":[{"term_id":"R-HSA-5653656","term_label":"Vesicle-mediated transport","supporting_discovery_ids":[0,5]},{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[6]}],"complexes":["MICAL3/ELKS/Rab8A exocytic complex","MICAL3/CRMP2/Numb complex"],"partners":["RAB8A","ELKS","RAB1","RAB35","KIF23","CRMP2","NUMB","CHEK1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q7RTP6","full_name":"[F-actin]-monooxygenase MICAL3","aliases":["Molecule interacting with CasL protein 3","MICAL-3"],"length_aa":2002,"mass_kda":224.3,"function":"Monooxygenase that promotes depolymerization of F-actin by mediating oxidation of specific methionine residues on actin to form methionine-sulfoxide, resulting in actin filament disassembly and preventing repolymerization. In the absence of actin, it also functions as a NADPH oxidase producing H(2)O(2). Seems to act as Rab effector protein and plays a role in vesicle trafficking. Involved in exocytic vesicles tethering and fusion: the monooxygenase activity is required for this process and implicates RAB8A associated with exocytotic vesicles. Required for cytokinesis. Contributes to stabilization and/or maturation of the intercellular bridge independently of its monooxygenase activity. Promotes recruitment of Rab8 and ERC1 to the intercellular bridge, and together these proteins are proposed to function in timely abscission","subcellular_location":"Cytoplasm; Cytoplasm, cell cortex; Cytoplasm, cytoskeleton; Nucleus; Midbody; Cytoplasm, cytoskeleton, spindle; Cytoplasm, cytoskeleton, cilium basal body","url":"https://www.uniprot.org/uniprotkb/Q7RTP6/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/MICAL3","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":[{"gene":"CSNK2B","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/MICAL3","total_profiled":1310},"omim":[{"mim_id":"616205","title":"MICRO RNA 648; MIR648","url":"https://www.omim.org/entry/616205"},{"mim_id":"608882","title":"MICROTUBULE-ASSOCIATED MONOOXYGENASE, CALPONIN AND LIM DOMAINS-CONTAINING, 3; MICAL3","url":"https://www.omim.org/entry/608882"},{"mim_id":"608881","title":"MICROTUBULE-ASSOCIATED MONOOXYGENASE, CALPONIN AND LIM DOMAINS-CONTAINING, 2; MICAL2","url":"https://www.omim.org/entry/608881"},{"mim_id":"601121","title":"PLACENTAL GROWTH FACTOR; PGF","url":"https://www.omim.org/entry/601121"},{"mim_id":"167414","title":"PAIRED BOX GENE 5; PAX5","url":"https://www.omim.org/entry/167414"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Nucleoplasm","reliability":"Supported"},{"location":"Midbody ring","reliability":"Supported"},{"location":"Plasma membrane","reliability":"Additional"},{"location":"Cytosol","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/MICAL3"},"hgnc":{"alias_symbol":["KIAA0819"],"prev_symbol":[]},"alphafold":{"accession":"Q7RTP6","domains":[{"cath_id":"3.50.50.60","chopping":"12-123_130-236_379-490","consensus_level":"high","plddt":94.5452,"start":12,"end":490},{"cath_id":"1.10.418.10","chopping":"519-627","consensus_level":"high","plddt":88.9955,"start":519,"end":627},{"cath_id":"2.10.110.10","chopping":"762-826","consensus_level":"medium","plddt":79.4485,"start":762,"end":826},{"cath_id":"3.30.70","chopping":"241-372","consensus_level":"medium","plddt":91.8045,"start":241,"end":372},{"cath_id":"1.20.1270","chopping":"1832-2002","consensus_level":"medium","plddt":82.086,"start":1832,"end":2002}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q7RTP6","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q7RTP6-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q7RTP6-F1-predicted_aligned_error_v6.png","plddt_mean":58.22},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=MICAL3","jax_strain_url":"https://www.jax.org/strain/search?query=MICAL3"},"sequence":{"accession":"Q7RTP6","fasta_url":"https://rest.uniprot.org/uniprotkb/Q7RTP6.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q7RTP6/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q7RTP6"}},"corpus_meta":[{"pmid":"22001757","id":"PMC_22001757","title":"Genome-wide association study identifies loci influencing concentrations of liver enzymes in plasma.","date":"2011","source":"Nature genetics","url":"https://pubmed.ncbi.nlm.nih.gov/22001757","citation_count":456,"is_preprint":false},{"pmid":"10048485","id":"PMC_10048485","title":"Prediction of the coding sequences of unidentified human genes. XII. The complete sequences of 100 new cDNA clones from brain which code for large proteins in vitro.","date":"1998","source":"DNA research : an international journal for rapid publication of reports on genes and genomes","url":"https://pubmed.ncbi.nlm.nih.gov/10048485","citation_count":190,"is_preprint":false},{"pmid":"21596566","id":"PMC_21596566","title":"Rab6, Rab8, and MICAL3 cooperate in controlling docking and fusion of exocytotic carriers.","date":"2011","source":"Current biology : CB","url":"https://pubmed.ncbi.nlm.nih.gov/21596566","citation_count":163,"is_preprint":false},{"pmid":"17971221","id":"PMC_17971221","title":"Boundary cap cells constrain spinal motor neuron somal migration at motor exit points by a semaphorin-plexin mechanism.","date":"2007","source":"Neural development","url":"https://pubmed.ncbi.nlm.nih.gov/17971221","citation_count":102,"is_preprint":false},{"pmid":"22307087","id":"PMC_22307087","title":"Reversible phosphocholination of Rab proteins by Legionella pneumophila effector proteins.","date":"2012","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/22307087","citation_count":91,"is_preprint":false},{"pmid":"22331357","id":"PMC_22331357","title":"Differential regulation of actin microfilaments by human MICAL proteins.","date":"2012","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/22331357","citation_count":85,"is_preprint":false},{"pmid":"26485645","id":"PMC_26485645","title":"The Ciliopathy Protein CC2D2A Associates with NINL and Functions in RAB8-MICAL3-Regulated Vesicle Trafficking.","date":"2015","source":"PLoS genetics","url":"https://pubmed.ncbi.nlm.nih.gov/26485645","citation_count":71,"is_preprint":false},{"pmid":"28373242","id":"PMC_28373242","title":"Emerging roles of MICAL family proteins - from actin oxidation to membrane trafficking during cytokinesis.","date":"2017","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/28373242","citation_count":69,"is_preprint":false},{"pmid":"15694364","id":"PMC_15694364","title":"The MICAL proteins and rab1: a possible link to the cytoskeleton?","date":"2005","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/15694364","citation_count":58,"is_preprint":false},{"pmid":"19223608","id":"PMC_19223608","title":"Muscle genome-wide expression profiling during disease evolution in mdx mice.","date":"2009","source":"Physiological genomics","url":"https://pubmed.ncbi.nlm.nih.gov/19223608","citation_count":54,"is_preprint":false},{"pmid":"30587593","id":"PMC_30587593","title":"Semaphorin signaling via MICAL3 induces symmetric cell division to expand breast cancer stem-like cells.","date":"2018","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/30587593","citation_count":49,"is_preprint":false},{"pmid":"29343822","id":"PMC_29343822","title":"The MICALs are a Family of F-actin Dismantling Oxidoreductases Conserved from Drosophila to Humans.","date":"2018","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/29343822","citation_count":40,"is_preprint":false},{"pmid":"36875759","id":"PMC_36875759","title":"MICAL-mediated oxidation of actin and its effects on cytoskeletal and cellular dynamics.","date":"2023","source":"Frontiers in cell and developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/36875759","citation_count":28,"is_preprint":false},{"pmid":"27528609","id":"PMC_27528609","title":"MICAL3 Flavoprotein Monooxygenase Forms a Complex with Centralspindlin and Regulates Cytokinesis.","date":"2016","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/27528609","citation_count":24,"is_preprint":false},{"pmid":"29044157","id":"PMC_29044157","title":"Facilitating identification of minimal protein binding domains by cross-linking mass spectrometry.","date":"2017","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/29044157","citation_count":18,"is_preprint":false},{"pmid":"34175685","id":"PMC_34175685","title":"Down-regulation of a panel of immune-related lncRNAs in breast cancer.","date":"2021","source":"Pathology, research and practice","url":"https://pubmed.ncbi.nlm.nih.gov/34175685","citation_count":14,"is_preprint":false},{"pmid":"25403488","id":"PMC_25403488","title":"MicroRNA 648 Targets ET-1 mRNA and is cotranscriptionally regulated with MICAL3 by PAX5.","date":"2014","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/25403488","citation_count":13,"is_preprint":false},{"pmid":"37725655","id":"PMC_37725655","title":"Disassembly of bundled F-actin and cellular remodeling via an interplay of Mical, cofilin, and F-actin crosslinkers.","date":"2023","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/37725655","citation_count":13,"is_preprint":false},{"pmid":"31949908","id":"PMC_31949908","title":"Structural and kinetic insights into flavin-containing monooxygenase and calponin-homology domains in human MICAL3.","date":"2020","source":"IUCrJ","url":"https://pubmed.ncbi.nlm.nih.gov/31949908","citation_count":12,"is_preprint":false},{"pmid":"22385522","id":"PMC_22385522","title":"Allelic expression analysis of the osteoarthritis susceptibility locus that maps to MICAL3.","date":"2012","source":"BMC medical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/22385522","citation_count":6,"is_preprint":false},{"pmid":"36140838","id":"PMC_36140838","title":"Blood Transcriptome Analysis of Beef Cow with Different Parity Revealed Candidate Genes and Gene Networks Regulating the Postpartum Diseases.","date":"2022","source":"Genes","url":"https://pubmed.ncbi.nlm.nih.gov/36140838","citation_count":5,"is_preprint":false},{"pmid":"33671465","id":"PMC_33671465","title":"Enhanced Production of the Mical Redox Domain for Enzymology and F-actin Disassembly Assays.","date":"2021","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/33671465","citation_count":3,"is_preprint":false},{"pmid":"39358552","id":"PMC_39358552","title":"CHK1 controls zygote pronuclear envelope breakdown by regulating F-actin through interacting with MICAL3.","date":"2024","source":"EMBO reports","url":"https://pubmed.ncbi.nlm.nih.gov/39358552","citation_count":2,"is_preprint":false},{"pmid":"38496508","id":"PMC_38496508","title":"Deep sequencing of proteotoxicity modifier genes uncovers a Presenilin-2/beta-amyloid-actin genetic risk module shared among alpha-synucleinopathies.","date":"2024","source":"bioRxiv : the preprint server for biology","url":"https://pubmed.ncbi.nlm.nih.gov/38496508","citation_count":1,"is_preprint":false},{"pmid":"33819263","id":"PMC_33819263","title":"ORN: Inferring patient-specific dysregulation status of pathway modules in cancer with OR-gate Network.","date":"2021","source":"PLoS computational biology","url":"https://pubmed.ncbi.nlm.nih.gov/33819263","citation_count":1,"is_preprint":false},{"pmid":"40397357","id":"PMC_40397357","title":"The Role of HbA1c in Parkinson's Disease: An Integrative Analysis by Single-Cell, Bulk Transcriptome and Mendelian Randomization.","date":"2025","source":"Molecular neurobiology","url":"https://pubmed.ncbi.nlm.nih.gov/40397357","citation_count":0,"is_preprint":false},{"pmid":"41795302","id":"PMC_41795302","title":"Mechanisms of heat and hypoxia defense in the sea cucumber Apostichopus japonicus: Insights from ubiquitination regulation.","date":"2026","source":"Comparative biochemistry and physiology. Part D, Genomics & proteomics","url":"https://pubmed.ncbi.nlm.nih.gov/41795302","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":15492,"output_tokens":3048,"usd":0.046098,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":10456,"output_tokens":3747,"usd":0.072978,"stage2_stop_reason":"end_turn"},"total_usd":0.119076,"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\": 2011,\n      \"finding\": \"MICAL3 interacts with both Rab8A and ELKS, linking these two proteins. Rab8A associates with exocytotic vesicles in a Rab6-dependent manner, and MICAL3 acts as a scaffold connecting Rab8A and ELKS at the cell cortex. Expression of a MICAL3 mutant with an inactive monooxygenase domain caused strong accumulation of secretory vesicles docked at the cell cortex that failed to fuse with the plasma membrane, indicating that MICAL3's monooxygenase activity is required for vesicle-docking complex remodeling and fusion.\",\n      \"method\": \"Co-immunoprecipitation, live-cell imaging of vesicle dynamics, dominant-negative monooxygenase mutant expression, FRAP\",\n      \"journal\": \"Current biology : CB\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, dominant-negative mutant with specific phenotypic readout, live imaging; replicated by subsequent studies\",\n      \"pmids\": [\"21596566\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"MICAL3 interacts with Rab1 in yeast two-hybrid and GST pulldown experiments. MICAL3 localizes to a microtubule-associated compartment, as treatment with the microtubule-depolymerizing drug nocodazole disrupts this localization, indicating a link between MICAL3 and the microtubule cytoskeleton.\",\n      \"method\": \"Yeast two-hybrid, GST pulldown, immunofluorescence with nocodazole treatment\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — yeast two-hybrid and GST pulldown for interaction, single lab, drug perturbation for localization\",\n      \"pmids\": [\"15694364\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"Knockdown of MICAL3 in chick motor neurons via RNAi caused ectopic positioning of motor neuron cell bodies in the peripheral nervous system, placing MICAL3 downstream of Neuropilin-2/Plexin-A2 semaphorin signaling as a cytoplasmic effector that controls somal positioning through cytoskeletal reorganization.\",\n      \"method\": \"RNA interference in chick embryo, analysis of motor neuron positioning\",\n      \"journal\": \"Neural development\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function RNAi with specific cellular phenotype, epistasis placement downstream of semaphorin/plexin, single lab\",\n      \"pmids\": [\"17971221\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"MICAL3 (as MICAL-3) was shown to interact with Rab1 and Rab35 effector binding is only slightly inhibited by phosphocholination, in contrast to GEF and GDI interactions which are strongly inhibited, demonstrating that MICAL-3 functions as a Rab effector protein.\",\n      \"method\": \"Biochemical binding assays with phosphocholinated Rab proteins, quantitative interaction measurements\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vitro biochemical assay with modified Rab proteins, single lab, defines MICAL3 as Rab effector\",\n      \"pmids\": [\"22307087\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Human MICAL3, like MICAL1 and MICAL2, is required for normal actin stress fiber organization in non-neural cells, and this actin-regulatory function depends on its generation of reactive oxygen species (ROS) via the monooxygenase domain.\",\n      \"method\": \"siRNA knockdown, ROS inhibitor treatment, immunofluorescence of actin stress fibers\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function with specific cytoskeletal phenotype and ROS dependency demonstrated, single lab\",\n      \"pmids\": [\"22331357\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"MICAL3 was identified as part of a NINL-associated interactome at the base of cilia, and interacts with Rab8 to regulate vesicle docking and fusion for cilia-directed cargo delivery. Genetic interaction between NINL and CC2D2A was demonstrated in zebrafish, and MICAL3 was linked to this pathway.\",\n      \"method\": \"Co-immunoprecipitation/interactome analysis, zebrafish morpholino knockdown, immunolocalization\",\n      \"journal\": \"PLoS genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP interactome, in vivo genetic interaction in zebrafish, localization data; single lab\",\n      \"pmids\": [\"26485645\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"MICAL3 forms a direct complex with the centralspindlin component MKLP1, is recruited to the central spindle and midbody during cytokinesis, and targets ELKS and Rab8A-positive vesicles to the midbody. Knockout of MICAL3 increases cytokinetic failure frequency and delays abscission. This scaffolding function is independent of MICAL3's enzymatic monooxygenase activity.\",\n      \"method\": \"Cross-linking mass spectrometry, MICAL3 knockout, cell biological assays (cytokinesis failure quantification, abscission timing), immunofluorescence\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — cross-linking MS for direct interaction, knockout with defined cellular phenotype, mechanistic dissection of enzymatic vs. scaffolding function; multiple orthogonal methods in one study\",\n      \"pmids\": [\"27528609\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Cross-linking mass spectrometry combined with deletion analysis defined the minimal binding domains of the MICAL3/ELKS/Rab8A complex involved in exocytosis, providing structural constraints on this protein assembly.\",\n      \"method\": \"Cross-linking mass spectrometry (XL-MS), deletion analysis, biochemical interaction assays\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — XL-MS with deletion validation defines binding interfaces; single lab, proof-of-concept study\",\n      \"pmids\": [\"29044157\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Human MICAL3 directly associates with F-actin, which activates its catalytic activity. MICAL3 uses NADPH-dependent redox activity to oxidize actin methionine residues M44/M47, dismantling filaments and limiting polymerization. MsrB/SelR reductase enzymes counteract MICAL3's effect on F-actin both in vitro and in vivo.\",\n      \"method\": \"In vitro F-actin binding and disassembly assays, NADPH consumption assays, genetic experiments in Drosophila, mass spectrometry of oxidized actin residues\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro reconstitution with purified proteins, site-specific modification identified by MS, genetic validation in vivo, reversibility demonstrated; multiple orthogonal methods\",\n      \"pmids\": [\"29343822\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Semaphorin 3 stimulation induces interaction among MICAL3, CRMP2, and Numb in breast cancer stem-like cells. MICAL3 monooxygenase activity (MO activity) is required for this interaction, for Numb protein accumulation, and for symmetric cell division. MICAL3 knockdown decreases sphere formation and shifts CSCs from symmetric to asymmetric division.\",\n      \"method\": \"Co-immunoprecipitation, MICAL3 siRNA knockdown, sphere formation assay, division mode quantification (symmetric vs. asymmetric)\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, loss-of-function with quantitative division mode phenotype, enzymatic activity requirement tested; single lab\",\n      \"pmids\": [\"30587593\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Crystal structure of human MICAL3 FMO and calponin-homology (CH) domains was solved. MICAL3 contains an FAD/NADP-binding Rossmann-fold domain for monooxygenase activity. Kinetic analysis showed that F-actin dramatically increases MICAL3 catalytic efficiency only when the CH domain is present. Mutation of Glu213 in the FMO domain and Arg530 in the CH domain abolished this F-actin-stimulated catalytic activation.\",\n      \"method\": \"X-ray crystallography, kinetic enzyme assays, site-directed mutagenesis (E213A, R530A)\",\n      \"journal\": \"IUCrJ\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structure plus kinetic assays plus mutagenesis in single study with multiple orthogonal methods; definitively maps F-actin-dependent activation mechanism\",\n      \"pmids\": [\"31949908\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"CHK1 physically interacts with MICAL3 in mouse zygotes, as identified by co-immunoprecipitation and mass spectrometry. Gain-of-function CHK1 mutants enhance interaction with MICAL3 and increase MICAL3 enzymatic activity, causing excessive F-actin depolymerization that disrupts pronuclear envelope breakdown.\",\n      \"method\": \"Co-immunoprecipitation and mass spectrometry of ~6000 mouse zygotes, gain-of-function CHK1 mutant analysis, F-actin imaging\",\n      \"journal\": \"EMBO reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP/MS interaction, gain-of-function mutant with specific cellular phenotype; single lab\",\n      \"pmids\": [\"39358552\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"MICAL3 is a flavoprotein monooxygenase that uses NADPH-dependent redox activity to oxidize actin methionine residues M44/M47, directly dismantling F-actin filaments in a process stimulated by F-actin binding to its calponin-homology domain (with Glu213 and Arg530 being critical for this activation); it also functions as a scaffold at the midbody and exocytotic sites by directly binding centralspindlin component MKLP1 and linking Rab8A-positive vesicles to ELKS, thereby coordinating vesicle docking/fusion during exocytosis and cytokinetic abscission, while additionally acting downstream of semaphorin/plexin signaling to regulate cytoskeletal reorganization, symmetric cell division, and pronuclear envelope breakdown through interactions with CRMP2/Numb and CHK1 respectively.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"MICAL3 is a flavoprotein monooxygenase that couples NADPH-dependent redox chemistry to direct remodeling of the actin cytoskeleton and to scaffolding of membrane-trafficking machinery [#8, #0]. As an enzyme, it binds F-actin through its calponin-homology (CH) domain, which dramatically stimulates its catalytic efficiency and allows it to oxidize actin methionine residues M44/M47, dismantling filaments and limiting polymerization; this oxidation is reversed by MsrB/SelR reductases [#8]. A crystal structure of the FMO and CH domains shows an FAD/NADP-binding Rossmann fold, and mutation of Glu213 (FMO) or Arg530 (CH) abolishes the F-actin-stimulated activation, defining the structural basis of substrate-coupled catalysis [#10]. Independently of its enzymatic activity, MICAL3 acts as a Rab effector and trafficking scaffold: it links Rab8A-positive exocytotic vesicles to ELKS at the cell cortex, where its monooxygenase activity is required for remodeling the docking complex to permit vesicle fusion [#0, #3]. The same scaffolding logic operates during cytokinesis, where MICAL3 binds the centralspindlin component MKLP1, is recruited to the central spindle and midbody, and targets ELKS and Rab8A vesicles there to support abscission [#6]. Through its actin-regulatory and trafficking functions MICAL3 contributes to broader cellular processes including semaphorin/plexin-directed cytoskeletal reorganization and motor neuron somal positioning [#2], symmetric division of cancer stem-like cells via a Semaphorin 3-induced MICAL3/CRMP2/Numb complex [#9], cilia-directed cargo delivery [#5], and pronuclear envelope breakdown downstream of CHK1 [#11].\",\n  \"teleology\": [\n    {\n      \"year\": 2005,\n      \"claim\": \"Established the first molecular handle on MICAL3 by placing it at the microtubule cytoskeleton as a candidate Rab1-binding protein, raising the question of whether it functions in membrane trafficking.\",\n      \"evidence\": \"Yeast two-hybrid and GST pulldown for Rab1 binding, immunofluorescence with nocodazole\",\n      \"pmids\": [\"15694364\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Rab1 interaction not validated by reciprocal endogenous Co-IP\", \"no functional consequence of the microtubule association defined\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Showed MICAL3 acts as a cytoplasmic effector of semaphorin signaling, answering whether it transduces guidance cues into cytoskeletal control in vivo.\",\n      \"evidence\": \"RNAi knockdown in chick embryo motor neurons, somal positioning analysis\",\n      \"pmids\": [\"17971221\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"molecular link between Neuropilin-2/Plexin-A2 and MICAL3 not biochemically defined\", \"single in vivo system\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Defined MICAL3 as a scaffold bridging Rab8A vesicles and ELKS at the cortex and showed its monooxygenase activity is required for vesicle fusion, establishing a catalytic role in exocytosis.\",\n      \"evidence\": \"Reciprocal Co-IP, live-cell vesicle imaging, dominant-negative monooxygenase mutant, FRAP\",\n      \"pmids\": [\"21596566\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"redox substrate at the docking site not identified\", \"structural geometry of the Rab8A-MICAL3-ELKS bridge unresolved\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Distinguished MICAL3 as a bona fide Rab effector and tied its monooxygenase-dependent ROS production to actin stress fiber organization in non-neural cells.\",\n      \"evidence\": \"Binding assays with phosphocholinated Rabs; siRNA knockdown with ROS inhibitor and actin immunofluorescence\",\n      \"pmids\": [\"22307087\", \"22331357\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"effector-versus-GEF/GDI selectivity tested only biochemically\", \"ROS-dependence inferred from inhibitors rather than direct substrate identification\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Extended MICAL3's Rab8-coupled trafficking role to the ciliary base, linking it to NINL-directed cargo delivery.\",\n      \"evidence\": \"Co-IP interactome, zebrafish morpholino knockdown, immunolocalization\",\n      \"pmids\": [\"26485645\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"direct versus indirect association with NINL not separated\", \"enzymatic requirement at cilia not tested\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Resolved that MICAL3's midbody function is structural by showing direct MKLP1 binding and vesicle targeting that supports abscission independently of monooxygenase activity, dissociating scaffolding from catalysis.\",\n      \"evidence\": \"Cross-linking mass spectrometry, MICAL3 knockout, cytokinesis-failure and abscission-timing assays\",\n      \"pmids\": [\"27528609\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"how MKLP1 recruitment is temporally regulated unknown\", \"whether catalytic activity contributes elsewhere in cytokinesis not addressed\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Defined the minimal binding interfaces of the MICAL3/ELKS/Rab8A exocytic complex, providing structural constraints on the assembly.\",\n      \"evidence\": \"Cross-linking mass spectrometry with deletion analysis\",\n      \"pmids\": [\"29044157\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"interfaces mapped by XL-MS, not high-resolution structure\", \"stoichiometry of the assembly not determined\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Established the core enzymatic mechanism: MICAL3 directly binds and oxidizes F-actin at methionine M44/M47 to disassemble filaments, a modification reversed by MsrB/SelR reductases, defining a redox switch on actin.\",\n      \"evidence\": \"In vitro F-actin binding/disassembly and NADPH assays, MS of oxidized residues, Drosophila genetics\",\n      \"pmids\": [\"29343822\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"regulation of catalysis in the cellular context not fully defined\", \"relationship between actin oxidation and vesicle-docking remodeling unresolved\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Connected MICAL3 monooxygenase activity to cell-fate control by showing a Semaphorin 3-induced MICAL3/CRMP2/Numb complex drives symmetric division of cancer stem-like cells.\",\n      \"evidence\": \"Co-IP, siRNA knockdown, sphere formation and symmetric/asymmetric division quantification\",\n      \"pmids\": [\"30587593\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"mechanism linking Numb stabilization to MICAL3 catalysis unclear\", \"directness of CRMP2/Numb interactions not established\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Provided the structural and kinetic basis for substrate-coupled activation, showing F-actin stimulates catalysis only when the CH domain is present and identifying Glu213 and Arg530 as essential for this allostery.\",\n      \"evidence\": \"X-ray crystallography of FMO/CH domains, kinetic assays, E213A/R530A mutagenesis\",\n      \"pmids\": [\"31949908\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"full-length enzyme structure including regulatory regions not solved\", \"conformational coupling between CH and FMO domains not visualized\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Identified CHK1 as an upstream regulator of MICAL3 activity in zygotes, where excessive MICAL3-driven actin depolymerization disrupts pronuclear envelope breakdown.\",\n      \"evidence\": \"Co-IP/MS from mouse zygotes, gain-of-function CHK1 mutants, F-actin imaging\",\n      \"pmids\": [\"39358552\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"whether CHK1 directly phosphorylates MICAL3 not shown\", \"single in vivo system, no reconstitution of the regulatory step\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How MICAL3's two activities — actin-oxidizing catalysis and Rab/centralspindlin scaffolding — are spatially and temporally integrated within a single cell remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"no unified model coupling redox actin remodeling to vesicle docking at exocytic and midbody sites\", \"upstream signals selecting between catalytic and scaffolding modes not defined\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0016491\", \"supporting_discovery_ids\": [4, 8, 10]},\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [8]},\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [8, 10]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 6]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [1, 8]},\n      {\"term_id\": \"GO:0005815\", \"supporting_discovery_ids\": [6]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-5653656\", \"supporting_discovery_ids\": [0, 5]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [6]}\n    ],\n    \"complexes\": [\n      \"MICAL3/ELKS/Rab8A exocytic complex\",\n      \"MICAL3/CRMP2/Numb complex\"\n    ],\n    \"partners\": [\n      \"RAB8A\",\n      \"ELKS\",\n      \"RAB1\",\n      \"RAB35\",\n      \"KIF23\",\n      \"CRMP2\",\n      \"NUMB\",\n      \"CHEK1\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}