{"gene":"MICALL1","run_date":"2026-06-10T02:59:50","timeline":{"discoveries":[{"year":2009,"finding":"MICAL-L1 directly interacts with EHD1 and Rab8a, colocalizing on tubular recycling endosomes. The MICAL-L1 C-terminal coiled-coil region is necessary and sufficient for localization to tubular membranes. MICAL-L1 depletion causes loss of EHD1-Rab8a interaction and absence of both proteins from tubular membranes, implicating MICAL-L1 as a scaffold/Rab effector that recruits and links EHD1 and Rab8a to tubular recycling endosomes to facilitate receptor recycling.","method":"Co-immunoprecipitation, live-cell imaging, siRNA knockdown, domain truncation experiments (coiled-coil region), transferrin and integrin receptor recycling assays","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal co-IP, domain mapping, live-cell imaging, functional recycling assay, replicated across multiple cell lines","pmids":["19864458"],"is_preprint":false},{"year":2011,"finding":"MICAL-L1 interacts with Rab35 and Arf6 in vivo. Active Rab35 overexpression impairs MICAL-L1 recruitment to tubular recycling endosomes, while Rab35 depletion enhances MICAL-L1 localization there. Arf6 forms a complex with MICAL-L1 and contributes to its endosomal recruitment. This positions Rab35 as a critical upstream regulator of MICAL-L1 and Arf6, with MICAL-L1 and Arf6 acting upstream of Rab8a.","method":"Co-immunoprecipitation, siRNA knockdown, overexpression of GTPase mutants (active/inactive), fluorescence microscopy","journal":"Traffic (Copenhagen, Denmark)","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal co-IP, epistasis via GTPase mutants and knockdown, two orthogonal methods, single lab","pmids":["21951725"],"is_preprint":false},{"year":2013,"finding":"MICAL-L1 and the BAR-domain protein syndapin2 both bind phosphatidic acid (a novel lipid component of recycling endosomes). Direct protein-protein interaction between MICAL-L1 and syndapin2 stabilizes membrane association. Phosphatidic acid in liposomes enhances syndapin2-mediated membrane tubulation in vitro, supporting a model in which MICAL-L1 and syndapin2 cooperate to nucleate tubular recycling endosome biogenesis.","method":"Lipid-binding assays (phosphatidic acid), in vitro liposome tubulation assay with recombinant syndapin2, co-immunoprecipitation, siRNA knockdown","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — in vitro reconstitution of tubulation, lipid-binding assays, co-IP, multiple orthogonal methods in single lab","pmids":["23596323"],"is_preprint":false},{"year":2014,"finding":"During NGF-induced neurite outgrowth, Rab35 recruits MICAL-L1 to Arf6-positive recycling endosomes, and MICAL-L1 in turn scaffolds Rab8, Rab13, and Rab36 to those endosomes. Knockdown of Rab36 impaired recruitment of the Rab36 effector JIP4 to recycling endosomes and inhibited neurite outgrowth without affecting Rab8 or Rab13 localization, demonstrating non-redundant downstream Rab functions.","method":"siRNA knockdown of Rab35, MICAL-L1, Rab8, Rab13, Rab36; fluorescence microscopy; neurite outgrowth assays in NGF-treated PC12 cells","journal":"Biology open","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — epistasis via knockdown and localization assays, multiple Rab knockdowns, single lab","pmids":["25086062"],"is_preprint":false},{"year":2014,"finding":"GRAF1 forms a complex with MICAL-L1 and EHD1 at tubular recycling endosomes. GRAF1 overexpression causes vesiculation of MICAL-L1-positive tubules; GRAF1 depletion impairs tubular recycling endosome vesiculation and delays receptor recycling. Co-addition of purified EHD1 and GRAF1 in a semi-permeabilized cell assay produces synergistic tubule vesiculation.","method":"Co-immunoprecipitation, siRNA knockdown, overexpression, semi-permeabilized cell vesiculation assay with purified proteins","journal":"Frontiers in cell and developmental biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP, functional vesiculation assay with purified proteins, knockdown phenotype, single lab","pmids":["25364729"],"is_preprint":false},{"year":2014,"finding":"MICAL-L1 depletion results in increased binucleated cells due to impaired recycling endosome transport during late cytokinesis, and also causes aberrant chromosome alignment and lagging chromosomes independently of EHD1, indicating an EHD1-independent role for MICAL-L1 earlier in mitosis. Both MICAL-L1 and EHD1 differentially influence microtubule dynamics during early and late mitosis.","method":"siRNA knockdown of MICAL-L1 and EHD1, binucleation assays, chromosome alignment imaging, microtubule dynamics analysis","journal":"Traffic (Copenhagen, Denmark)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — epistasis via differential knockdown phenotypes, multiple mitotic readouts, single lab","pmids":["25287187"],"is_preprint":false},{"year":2014,"finding":"MICAL-L1 is required for growth-factor- and integrin-induced Src activation and transport to the cell periphery. MICAL-L1 depletion impairs focal adhesion turnover, cell spreading, and migration. MICAL-L1-mediated recruitment of EHD1 to Src-containing recycling endosomes is required for Src release from the perinuclear recycling compartment in response to growth factor stimulation.","method":"siRNA knockdown of MICAL-L1 and EHD1, co-localization by fluorescence microscopy, Src activation assays, focal adhesion turnover assays, migration assays in HeLa cells and human fibroblasts","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — epistasis via knockdown, multiple functional readouts, single lab","pmids":["24481818"],"is_preprint":false},{"year":2017,"finding":"MICALL1 is a transcriptional target of p53; functional p53-binding motifs were identified ~3000 bp upstream of the MICALL1 gene. In response to DNA damage, MICALL1 co-localizes with RAB8A and CD2AP at tubular recycling endosomes; siRNA depletion of p53 or MICALL1 prevents this localization, demonstrating that p53 regulates tubular recycling endosome biogenesis via MICALL1.","method":"Multi-omics (mass spectrometry, cDNA microarray), p53 binding motif identification, siRNA knockdown, co-localization by fluorescence microscopy","journal":"International journal of oncology","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — p53-binding motif identified, siRNA epistasis, localization assay, multiple methods but single lab","pmids":["28714518"],"is_preprint":false},{"year":2019,"finding":"MICAL-L1 localizes to cilia and centrosomes (including non-ciliated cells). MICAL-L1 knockdown impairs ciliogenesis similarly to EHD1 knockdown, prevents CP110 removal from the mother centriole, and causes EHD1 to fail to localize to basal bodies. Mass spectrometry and direct binding assays identified α-tubulin–β-tubulin heterodimers and γ-tubulin as direct MICAL-L1 interaction partners, supporting a model in which centriolar tubulins anchor MICAL-L1 to recruit EHD1 for ciliogenesis.","method":"siRNA knockdown, fluorescence microscopy (localization to cilia/centrosomes), mass spectrometry, direct protein-binding assays (tubulin interactions)","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct binding assay + knockdown with defined ciliogenesis phenotypes, single lab, two orthogonal methods","pmids":["31615969"],"is_preprint":false},{"year":2021,"finding":"PACSIN2 recruits EHD4 and MICAL-L1 to the rear end of asymmetric adherens junctions in endothelial cells, forming a recycling endosome-like tubular structure. This junctional PACSIN2/EHD4/MICAL-L1 complex controls local VE-cadherin trafficking and coordinates polarized endothelial migration and angiogenesis.","method":"Co-immunoprecipitation, fluorescence microscopy, siRNA/shRNA knockdown, VE-cadherin trafficking assays, angiogenesis sprouting assays","journal":"Nature communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP, localization, functional trafficking and angiogenesis assays, single lab","pmids":["33972531"],"is_preprint":false},{"year":2021,"finding":"MICAL-L1 depletion impairs delivery of a subset of cargo proteins to the cell surface. The MICAL-L1 RBD domain contributes to PACSIN-mediated membrane tubulation in vitro. Two hydrophobic residues at the MICAL-L1 C-terminus are important for phosphatidic acid binding and for association with membrane tubules. MICAL-L1 associates with Golgi apparatus markers and recycling endosomes.","method":"shRNA knockdown, synchronized secretion assay, in vitro membrane tubulation assay with recombinant MICAL-L1-RBD, confocal and STORM super-resolution microscopy, site-directed mutagenesis of C-terminal hydrophobic residues","journal":"Biology open","confidence":"Medium","confidence_rationale":"Tier 1–2 / Moderate — in vitro tubulation, mutagenesis, functional secretion assay, single lab with multiple methods","pmids":["34100897"],"is_preprint":false},{"year":2024,"finding":"MICAL-L1 directly interacts with FCHSD2 and recruits it to the endosomal membrane. FCHSD2 recruitment by MICAL-L1 is required for ARP2/3-mediated branched actin generation, endosome fission, and receptor recycling. Because MICAL-L1 recruits FCHSD2 prior to EHD1, MICAL-L1 orchestrates the sequential steps of endosomal fission by bridging early actin-driven constriction (via FCHSD2/ARP2/3) and subsequent nucleotide hydrolysis/fission (via EHD1).","method":"Co-immunoprecipitation, siRNA knockdown of MICAL-L1 and FCHSD2, ARP2/3 actin branching assays, endosome fission assays, receptor recycling assays","journal":"Molecular biology of the cell","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP, knockdown with defined mechanistic phenotypes, actin branching assay, single lab","pmids":["39382837"],"is_preprint":false},{"year":2025,"finding":"CD2AP and CIN85 are novel constituents of tubular recycling endosomes recruited by MICAL-L1 via their SH3 domains. Depletion of either CD2AP or CIN85 impairs recycling endosome function. MICAL-L1 scaffolding of these proteins likely impacts recycling through effects on the actin cytoskeleton.","method":"Co-immunoprecipitation (SH3 domain interactions), siRNA knockdown of CD2AP and CIN85, receptor recycling assays, fluorescence microscopy","journal":"Traffic (Copenhagen, Denmark)","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — co-IP showing SH3-mediated interaction, knockdown with recycling phenotype, single lab","pmids":["40740057"],"is_preprint":false},{"year":2014,"finding":"MICAL-L1 is necessary for ETEN (elongated tubular endosomal network) remodeling originating from the endocytic recycling compartment in human dendritic cells, as demonstrated by knockdown experiments.","method":"siRNA knockdown, fluorescence microscopy","journal":"Communicative & integrative biology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single knockdown experiment, single localization readout, single lab","pmids":["26478765"],"is_preprint":false}],"current_model":"MICAL-L1 functions as a multi-valent endosomal scaffold protein on tubular recycling endosomes (TREs), where its C-terminal coiled-coil domain mediates membrane association via phosphatidic acid binding; it acts as a Rab35 effector that bridges upstream GTPases (Rab35, Arf6) with downstream effectors (Rab8a, Rab13, Rab36), recruits EHD1 and syndapin2 for TRE biogenesis, recruits FCHSD2/ARP2/3 for early actin-driven endosome fission followed by EHD1-mediated nucleotide hydrolysis fission, scaffolds CD2AP, CIN85, and GRAF1 for TRE vesiculation, coordinates Src and VE-cadherin trafficking to regulate cell migration and angiogenesis, and also localizes to centrosomes (anchored by tubulins) to recruit EHD1 for ciliogenesis."},"narrative":{"mechanistic_narrative":"MICALL1 (MICAL-L1) is a multivalent membrane scaffold that organizes tubular recycling endosomes (TREs) and coordinates receptor recycling, with downstream consequences for cell migration, mitosis, and ciliogenesis [PMID:19864458, PMID:21951725]. Its C-terminal coiled-coil region is necessary and sufficient for targeting to tubular membranes, and membrane association is reinforced by direct phosphatidic acid binding through C-terminal hydrophobic residues [PMID:19864458, PMID:23596323, PMID:34100897]. MICAL-L1 acts as a Rab35 effector that bridges upstream GTPases to downstream traffic: Rab35 and Arf6 govern its endosomal recruitment, after which MICAL-L1 scaffolds the non-redundant downstream Rabs Rab8a, Rab13, and Rab36 onto recycling endosomes [PMID:21951725, PMID:25086062]. On TREs it nucleates a fission and biogenesis machinery, partnering with the BAR-domain protein syndapin2 to drive membrane tubulation [PMID:23596323], and orchestrating sequential endosome fission by first recruiting FCHSD2 to trigger ARP2/3-mediated branched actin constriction and then EHD1 for nucleotide-hydrolysis-driven scission [PMID:19864458, PMID:39382837]. It additionally scaffolds GRAF1 and the SH3-domain adaptors CD2AP and CIN85 to promote tubule vesiculation and recycling [PMID:25364729, PMID:40740057]. Through this trafficking function MICAL-L1 supports growth-factor- and integrin-induced Src activation, focal adhesion turnover, and migration [PMID:24481818], and within endothelial junctions a PACSIN2/EHD4/MICAL-L1 complex directs VE-cadherin trafficking during angiogenesis [PMID:33972531]. Beyond recycling, MICAL-L1 binds α/β- and γ-tubulin to anchor at centrosomes where it recruits EHD1 to drive CP110 removal and ciliogenesis [PMID:31615969], and is required for recycling-endosome-dependent cytokinesis and chromosome alignment in mitosis [PMID:25287187]. MICALL1 is a transcriptional target of p53 that links DNA damage signaling to TRE biogenesis [PMID:28714518].","teleology":[{"year":2009,"claim":"Established MICAL-L1 as the scaffold that physically links EHD1 and Rab8a on tubular recycling endosomes, answering how these recycling factors are co-recruited to tubular membranes.","evidence":"Reciprocal co-IP, domain truncation mapping the coiled-coil, siRNA knockdown, and transferrin/integrin recycling assays across cell lines","pmids":["19864458"],"confidence":"High","gaps":["Did not define upstream GTPase control of MICAL-L1 recruitment","No structural detail of the coiled-coil membrane interaction"]},{"year":2011,"claim":"Placed MICAL-L1 within a GTPase hierarchy, showing Rab35 and Arf6 act upstream to control its endosomal recruitment and that MICAL-L1/Arf6 lie upstream of Rab8a.","evidence":"Co-IP, siRNA knockdown, and active/inactive GTPase mutant overexpression with fluorescence microscopy","pmids":["21951725"],"confidence":"High","gaps":["Mechanism by which active Rab35 displaces MICAL-L1 from TREs not resolved","Single lab"]},{"year":2013,"claim":"Defined the lipid basis of TRE membrane association, showing MICAL-L1 and syndapin2 bind phosphatidic acid and cooperate to nucleate tubulation.","evidence":"Lipid-binding assays, in vitro liposome tubulation with recombinant syndapin2, and co-IP","pmids":["23596323"],"confidence":"High","gaps":["In vivo contribution of phosphatidic acid to TRE biogenesis not quantified","Did not map the MICAL-L1 lipid-binding residues (later addressed in #10)"]},{"year":2014,"claim":"Showed MICAL-L1 scaffolds multiple non-redundant downstream Rabs (Rab8, Rab13, Rab36) during NGF-induced neurite outgrowth, with Rab36/JIP4 having a distinct functional output.","evidence":"siRNA knockdown of Rab35/MICAL-L1/Rab8/Rab13/Rab36 and neurite outgrowth assays in PC12 cells","pmids":["25086062"],"confidence":"Medium","gaps":["Direct binding of each Rab to MICAL-L1 not biochemically dissected","Generality beyond NGF/PC12 system unknown"]},{"year":2014,"claim":"Identified GRAF1 as a MICAL-L1/EHD1-associated factor driving TRE vesiculation, linking the scaffold to membrane scission.","evidence":"Co-IP, knockdown, and a semi-permeabilized cell vesiculation assay with purified EHD1 and GRAF1","pmids":["25364729"],"confidence":"Medium","gaps":["Order of GRAF1 versus other fission factors not established","Single lab"]},{"year":2014,"claim":"Connected MICAL-L1 trafficking function to Src activation, focal adhesion turnover, and migration, defining a physiological output of TRE recycling.","evidence":"siRNA knockdown of MICAL-L1/EHD1, Src activation, focal adhesion turnover, and migration assays in HeLa and fibroblasts","pmids":["24481818"],"confidence":"Medium","gaps":["Direct cargo-binding mechanism for Src vesicles not shown","Single lab"]},{"year":2014,"claim":"Revealed mitotic roles, including an EHD1-independent function in chromosome alignment and an EHD1-dependent recycling role in cytokinesis.","evidence":"Differential siRNA knockdown of MICAL-L1 and EHD1 with binucleation, chromosome alignment, and microtubule dynamics readouts","pmids":["25287187"],"confidence":"Medium","gaps":["Molecular partner for the EHD1-independent mitotic function unidentified","Mechanism linking MICAL-L1 to microtubule dynamics unclear"]},{"year":2017,"claim":"Identified MICALL1 as a direct p53 transcriptional target, linking DNA damage signaling to TRE biogenesis.","evidence":"Multi-omics, p53-binding motif identification ~3 kb upstream, siRNA depletion of p53/MICALL1, and co-localization with RAB8A/CD2AP","pmids":["28714518"],"confidence":"Medium","gaps":["Functional consequence of p53-driven TRE biogenesis for DNA damage outcomes not established","Single lab"]},{"year":2019,"claim":"Extended MICAL-L1 function to the centrosome/cilium, showing it binds tubulins to anchor at centrioles and recruit EHD1 for CP110 removal and ciliogenesis.","evidence":"siRNA knockdown with ciliogenesis phenotypes, mass spectrometry, and direct binding assays to α/β- and γ-tubulin","pmids":["31615969"],"confidence":"Medium","gaps":["How a recycling scaffold also acts at centrioles mechanistically distinct from TREs unclear","Single lab"]},{"year":2021,"claim":"Demonstrated a junctional PACSIN2/EHD4/MICAL-L1 complex controlling VE-cadherin trafficking, polarized endothelial migration, and angiogenesis.","evidence":"Co-IP, knockdown, VE-cadherin trafficking assays, and angiogenesis sprouting assays in endothelial cells","pmids":["33972531"],"confidence":"Medium","gaps":["Whether this is the same TRE machinery repurposed at junctions not fully resolved","Single lab"]},{"year":2021,"claim":"Mapped C-terminal hydrophobic residues required for phosphatidic acid binding and defined the RBD contribution to PACSIN-mediated tubulation and surface cargo delivery.","evidence":"shRNA knockdown, synchronized secretion assay, in vitro tubulation with recombinant MICAL-L1-RBD, STORM imaging, and site-directed mutagenesis","pmids":["34100897"],"confidence":"Medium","gaps":["Identity of the affected cargo subset incompletely defined","Golgi association mechanism not detailed"]},{"year":2024,"claim":"Resolved the sequential logic of endosome fission, showing MICAL-L1 recruits FCHSD2/ARP2/3 for early actin-driven constriction before EHD1-mediated scission.","evidence":"Co-IP, siRNA knockdown of MICAL-L1/FCHSD2, ARP2/3 actin branching, endosome fission, and recycling assays","pmids":["39382837"],"confidence":"Medium","gaps":["Temporal coordination measured indirectly","Single lab"]},{"year":2025,"claim":"Added CD2AP and CIN85 as SH3-domain-recruited TRE constituents acting through the actin cytoskeleton in recycling.","evidence":"Co-IP mapping SH3 interactions, siRNA knockdown of CD2AP/CIN85, and receptor recycling assays","pmids":["40740057"],"confidence":"Medium","gaps":["Direct actin-cytoskeleton link inferred, not demonstrated","Single lab"]},{"year":null,"claim":"How MICAL-L1's many partner interactions are spatially and temporally segregated across TREs, centrosomes, junctions, and the mitotic apparatus remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural model of the multivalent scaffold with bound partners","Regulation of context-specific complex assembly unknown","Whether disease-relevant phenotypes arise from MICALL1 loss not established"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,1,3,4,11,12]},{"term_id":"GO:0008289","term_label":"lipid binding","supporting_discovery_ids":[2,10]},{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[8]}],"localization":[{"term_id":"GO:0005768","term_label":"endosome","supporting_discovery_ids":[0,1,2,4,11,12]},{"term_id":"GO:0005815","term_label":"microtubule organizing center","supporting_discovery_ids":[8]},{"term_id":"GO:0005929","term_label":"cilium","supporting_discovery_ids":[8]},{"term_id":"GO:0005794","term_label":"Golgi apparatus","supporting_discovery_ids":[10]}],"pathway":[{"term_id":"R-HSA-5653656","term_label":"Vesicle-mediated transport","supporting_discovery_ids":[0,4,11,12]},{"term_id":"R-HSA-9609507","term_label":"Protein localization","supporting_discovery_ids":[6,9,10]},{"term_id":"R-HSA-1852241","term_label":"Organelle biogenesis and maintenance","supporting_discovery_ids":[8]}],"complexes":["MICAL-L1/EHD1/Rab8a TRE scaffold","PACSIN2/EHD4/MICAL-L1 junctional complex","MICAL-L1/EHD1/GRAF1 complex"],"partners":["EHD1","RAB8A","RAB35","ARF6","PACSIN2","FCHSD2","CD2AP","CIN85"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q8N3F8","full_name":"MICAL-like protein 1","aliases":["Molecule interacting with Rab13","MIRab13"],"length_aa":863,"mass_kda":93.4,"function":"Lipid-binding protein with higher affinity for phosphatidic acid, a lipid enriched in recycling endosome membranes. On endosome membranes, acts as a downstream effector of Rab proteins recruiting cytosolic proteins to regulate membrane tubulation (PubMed:19864458, PubMed:20801876, PubMed:23596323, PubMed:34100897). Involved in a late step of receptor-mediated endocytosis regulating for instance endocytosed-EGF receptor trafficking (PubMed:21795389). Alternatively, regulates slow endocytic recycling of endocytosed proteins back to the plasma membrane (PubMed:19864458). Also involved in cargo protein delivery to the plasma membrane (PubMed:34100897). Plays a role in ciliogenesis coordination, recruits EHD1 to primary cilium where it is anchored to the centriole through interaction with tubulins (PubMed:31615969). May indirectly play a role in neurite outgrowth (By similarity)","subcellular_location":"Recycling endosome membrane; Late endosome membrane; Cell projection, cilium membrane; Cytoplasm, cytoskeleton, microtubule organizing center, centrosome, centriole","url":"https://www.uniprot.org/uniprotkb/Q8N3F8/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/MICALL1","classification":"Not Classified","n_dependent_lines":20,"n_total_lines":1208,"dependency_fraction":0.016556291390728478},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"CAPZB","stoichiometry":0.2},{"gene":"PACSIN2","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/MICALL1","total_profiled":1310},"omim":[{"mim_id":"619563","title":"MICAL-LIKE PROTEIN 1; MICALL1","url":"https://www.omim.org/entry/619563"},{"mim_id":"605888","title":"EH DOMAIN-CONTAINING 1; EHD1","url":"https://www.omim.org/entry/605888"},{"mim_id":"190090","title":"SRC PROTOONCOGENE, NONRECEPTOR TYROSINE KINASE; SRC","url":"https://www.omim.org/entry/190090"},{"mim_id":"142460","title":"SYNDECAN 2; SDC2","url":"https://www.omim.org/entry/142460"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Cell Junctions","reliability":"Supported"},{"location":"Centriolar satellite","reliability":"Supported"},{"location":"Cytosol","reliability":"Supported"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in many","driving_tissues":[],"url":"https://www.proteinatlas.org/search/MICALL1"},"hgnc":{"alias_symbol":["MIRAB13","KIAA1668","MICAL-L1"],"prev_symbol":[]},"alphafold":{"accession":"Q8N3F8","domains":[{"cath_id":"1.10.418.10","chopping":"2-110","consensus_level":"medium","plddt":89.1156,"start":2,"end":110},{"cath_id":"2.10.110.10","chopping":"163-220","consensus_level":"medium","plddt":83.6881,"start":163,"end":220},{"cath_id":"-","chopping":"676-827","consensus_level":"medium","plddt":84.8248,"start":676,"end":827}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8N3F8","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q8N3F8-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q8N3F8-F1-predicted_aligned_error_v6.png","plddt_mean":57.88},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=MICALL1","jax_strain_url":"https://www.jax.org/strain/search?query=MICALL1"},"sequence":{"accession":"Q8N3F8","fasta_url":"https://rest.uniprot.org/uniprotkb/Q8N3F8.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q8N3F8/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8N3F8"}},"corpus_meta":[{"pmid":"19864458","id":"PMC_19864458","title":"MICAL-L1 links EHD1 to tubular recycling endosomes and regulates receptor recycling.","date":"2009","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/19864458","citation_count":151,"is_preprint":false},{"pmid":"21951725","id":"PMC_21951725","title":"MICAL-L1 is a tubular endosomal membrane hub that connects Rab35 and Arf6 with Rab8a.","date":"2011","source":"Traffic (Copenhagen, Denmark)","url":"https://pubmed.ncbi.nlm.nih.gov/21951725","citation_count":91,"is_preprint":false},{"pmid":"23596323","id":"PMC_23596323","title":"Cooperation of MICAL-L1, syndapin2, and phosphatidic acid in tubular recycling endosome biogenesis.","date":"2013","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/23596323","citation_count":89,"is_preprint":false},{"pmid":"25086062","id":"PMC_25086062","title":"Rab35 promotes the recruitment of Rab8, Rab13 and Rab36 to recycling endosomes through MICAL-L1 during neurite outgrowth.","date":"2014","source":"Biology open","url":"https://pubmed.ncbi.nlm.nih.gov/25086062","citation_count":71,"is_preprint":false},{"pmid":"25364729","id":"PMC_25364729","title":"GRAF1 forms a complex with MICAL-L1 and EHD1 to cooperate in tubular recycling endosome vesiculation.","date":"2014","source":"Frontiers in cell and developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/25364729","citation_count":37,"is_preprint":false},{"pmid":"33972531","id":"PMC_33972531","title":"A junctional PACSIN2/EHD4/MICAL-L1 complex coordinates VE-cadherin trafficking for endothelial migration and angiogenesis.","date":"2021","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/33972531","citation_count":34,"is_preprint":false},{"pmid":"31615969","id":"PMC_31615969","title":"MICAL-L1 coordinates ciliogenesis by recruiting EHD1 to the primary cilium.","date":"2019","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/31615969","citation_count":24,"is_preprint":false},{"pmid":"20585517","id":"PMC_20585517","title":"MICAL-L1: An unusual Rab effector that links EHD1 to tubular recycling endosomes.","date":"2010","source":"Communicative & integrative biology","url":"https://pubmed.ncbi.nlm.nih.gov/20585517","citation_count":23,"is_preprint":false},{"pmid":"25287187","id":"PMC_25287187","title":"Novel functions for the endocytic regulatory proteins MICAL-L1 and EHD1 in mitosis.","date":"2014","source":"Traffic (Copenhagen, Denmark)","url":"https://pubmed.ncbi.nlm.nih.gov/25287187","citation_count":22,"is_preprint":false},{"pmid":"23060965","id":"PMC_23060965","title":"Trafficking cascades mediated by Rab35 and its membrane hub effector, MICAL-L1.","date":"2012","source":"Communicative & integrative biology","url":"https://pubmed.ncbi.nlm.nih.gov/23060965","citation_count":18,"is_preprint":false},{"pmid":"24481818","id":"PMC_24481818","title":"Regulation of Src trafficking and activation by the endocytic regulatory proteins MICAL-L1 and EHD1.","date":"2014","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/24481818","citation_count":17,"is_preprint":false},{"pmid":"26478765","id":"PMC_26478765","title":"MICAL-L1-related and unrelated mechanisms underlying elongated tubular endosomal network (ETEN) in human dendritic cells.","date":"2014","source":"Communicative & integrative biology","url":"https://pubmed.ncbi.nlm.nih.gov/26478765","citation_count":10,"is_preprint":false},{"pmid":"28714518","id":"PMC_28714518","title":"Regulation of tubular recycling endosome biogenesis by the p53-MICALL1 pathway.","date":"2017","source":"International journal of oncology","url":"https://pubmed.ncbi.nlm.nih.gov/28714518","citation_count":8,"is_preprint":false},{"pmid":"34100897","id":"PMC_34100897","title":"MICAL-L1 is required for cargo protein delivery to the cell surface.","date":"2021","source":"Biology open","url":"https://pubmed.ncbi.nlm.nih.gov/34100897","citation_count":4,"is_preprint":false},{"pmid":"39382837","id":"PMC_39382837","title":"Endosomal actin branching, fission, and receptor recycling require FCHSD2 recruitment by MICAL-L1.","date":"2024","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/39382837","citation_count":4,"is_preprint":false},{"pmid":"40740057","id":"PMC_40740057","title":"CIN85 and CD2AP Are Novel Constituents of Dynamic Tubular Recycling Endosomes That Regulate Recycling Upon Recruitment by MICAL-L1.","date":"2025","source":"Traffic (Copenhagen, Denmark)","url":"https://pubmed.ncbi.nlm.nih.gov/40740057","citation_count":3,"is_preprint":false},{"pmid":"38979241","id":"PMC_38979241","title":"Endosomal actin branching, fission and receptor recycling require FCHSD2 recruitment by MICAL-L1.","date":"2024","source":"bioRxiv : the preprint server for biology","url":"https://pubmed.ncbi.nlm.nih.gov/38979241","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":9553,"output_tokens":4028,"usd":0.04454,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":11724,"output_tokens":4420,"usd":0.08456,"stage2_stop_reason":"end_turn"},"total_usd":0.1291,"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\": 2009,\n      \"finding\": \"MICAL-L1 directly interacts with EHD1 and Rab8a, colocalizing on tubular recycling endosomes. The MICAL-L1 C-terminal coiled-coil region is necessary and sufficient for localization to tubular membranes. MICAL-L1 depletion causes loss of EHD1-Rab8a interaction and absence of both proteins from tubular membranes, implicating MICAL-L1 as a scaffold/Rab effector that recruits and links EHD1 and Rab8a to tubular recycling endosomes to facilitate receptor recycling.\",\n      \"method\": \"Co-immunoprecipitation, live-cell imaging, siRNA knockdown, domain truncation experiments (coiled-coil region), transferrin and integrin receptor recycling assays\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal co-IP, domain mapping, live-cell imaging, functional recycling assay, replicated across multiple cell lines\",\n      \"pmids\": [\"19864458\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"MICAL-L1 interacts with Rab35 and Arf6 in vivo. Active Rab35 overexpression impairs MICAL-L1 recruitment to tubular recycling endosomes, while Rab35 depletion enhances MICAL-L1 localization there. Arf6 forms a complex with MICAL-L1 and contributes to its endosomal recruitment. This positions Rab35 as a critical upstream regulator of MICAL-L1 and Arf6, with MICAL-L1 and Arf6 acting upstream of Rab8a.\",\n      \"method\": \"Co-immunoprecipitation, siRNA knockdown, overexpression of GTPase mutants (active/inactive), fluorescence microscopy\",\n      \"journal\": \"Traffic (Copenhagen, Denmark)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal co-IP, epistasis via GTPase mutants and knockdown, two orthogonal methods, single lab\",\n      \"pmids\": [\"21951725\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"MICAL-L1 and the BAR-domain protein syndapin2 both bind phosphatidic acid (a novel lipid component of recycling endosomes). Direct protein-protein interaction between MICAL-L1 and syndapin2 stabilizes membrane association. Phosphatidic acid in liposomes enhances syndapin2-mediated membrane tubulation in vitro, supporting a model in which MICAL-L1 and syndapin2 cooperate to nucleate tubular recycling endosome biogenesis.\",\n      \"method\": \"Lipid-binding assays (phosphatidic acid), in vitro liposome tubulation assay with recombinant syndapin2, co-immunoprecipitation, siRNA knockdown\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — in vitro reconstitution of tubulation, lipid-binding assays, co-IP, multiple orthogonal methods in single lab\",\n      \"pmids\": [\"23596323\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"During NGF-induced neurite outgrowth, Rab35 recruits MICAL-L1 to Arf6-positive recycling endosomes, and MICAL-L1 in turn scaffolds Rab8, Rab13, and Rab36 to those endosomes. Knockdown of Rab36 impaired recruitment of the Rab36 effector JIP4 to recycling endosomes and inhibited neurite outgrowth without affecting Rab8 or Rab13 localization, demonstrating non-redundant downstream Rab functions.\",\n      \"method\": \"siRNA knockdown of Rab35, MICAL-L1, Rab8, Rab13, Rab36; fluorescence microscopy; neurite outgrowth assays in NGF-treated PC12 cells\",\n      \"journal\": \"Biology open\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — epistasis via knockdown and localization assays, multiple Rab knockdowns, single lab\",\n      \"pmids\": [\"25086062\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"GRAF1 forms a complex with MICAL-L1 and EHD1 at tubular recycling endosomes. GRAF1 overexpression causes vesiculation of MICAL-L1-positive tubules; GRAF1 depletion impairs tubular recycling endosome vesiculation and delays receptor recycling. Co-addition of purified EHD1 and GRAF1 in a semi-permeabilized cell assay produces synergistic tubule vesiculation.\",\n      \"method\": \"Co-immunoprecipitation, siRNA knockdown, overexpression, semi-permeabilized cell vesiculation assay with purified proteins\",\n      \"journal\": \"Frontiers in cell and developmental biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP, functional vesiculation assay with purified proteins, knockdown phenotype, single lab\",\n      \"pmids\": [\"25364729\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"MICAL-L1 depletion results in increased binucleated cells due to impaired recycling endosome transport during late cytokinesis, and also causes aberrant chromosome alignment and lagging chromosomes independently of EHD1, indicating an EHD1-independent role for MICAL-L1 earlier in mitosis. Both MICAL-L1 and EHD1 differentially influence microtubule dynamics during early and late mitosis.\",\n      \"method\": \"siRNA knockdown of MICAL-L1 and EHD1, binucleation assays, chromosome alignment imaging, microtubule dynamics analysis\",\n      \"journal\": \"Traffic (Copenhagen, Denmark)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — epistasis via differential knockdown phenotypes, multiple mitotic readouts, single lab\",\n      \"pmids\": [\"25287187\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"MICAL-L1 is required for growth-factor- and integrin-induced Src activation and transport to the cell periphery. MICAL-L1 depletion impairs focal adhesion turnover, cell spreading, and migration. MICAL-L1-mediated recruitment of EHD1 to Src-containing recycling endosomes is required for Src release from the perinuclear recycling compartment in response to growth factor stimulation.\",\n      \"method\": \"siRNA knockdown of MICAL-L1 and EHD1, co-localization by fluorescence microscopy, Src activation assays, focal adhesion turnover assays, migration assays in HeLa cells and human fibroblasts\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — epistasis via knockdown, multiple functional readouts, single lab\",\n      \"pmids\": [\"24481818\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"MICALL1 is a transcriptional target of p53; functional p53-binding motifs were identified ~3000 bp upstream of the MICALL1 gene. In response to DNA damage, MICALL1 co-localizes with RAB8A and CD2AP at tubular recycling endosomes; siRNA depletion of p53 or MICALL1 prevents this localization, demonstrating that p53 regulates tubular recycling endosome biogenesis via MICALL1.\",\n      \"method\": \"Multi-omics (mass spectrometry, cDNA microarray), p53 binding motif identification, siRNA knockdown, co-localization by fluorescence microscopy\",\n      \"journal\": \"International journal of oncology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — p53-binding motif identified, siRNA epistasis, localization assay, multiple methods but single lab\",\n      \"pmids\": [\"28714518\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"MICAL-L1 localizes to cilia and centrosomes (including non-ciliated cells). MICAL-L1 knockdown impairs ciliogenesis similarly to EHD1 knockdown, prevents CP110 removal from the mother centriole, and causes EHD1 to fail to localize to basal bodies. Mass spectrometry and direct binding assays identified α-tubulin–β-tubulin heterodimers and γ-tubulin as direct MICAL-L1 interaction partners, supporting a model in which centriolar tubulins anchor MICAL-L1 to recruit EHD1 for ciliogenesis.\",\n      \"method\": \"siRNA knockdown, fluorescence microscopy (localization to cilia/centrosomes), mass spectrometry, direct protein-binding assays (tubulin interactions)\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct binding assay + knockdown with defined ciliogenesis phenotypes, single lab, two orthogonal methods\",\n      \"pmids\": [\"31615969\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"PACSIN2 recruits EHD4 and MICAL-L1 to the rear end of asymmetric adherens junctions in endothelial cells, forming a recycling endosome-like tubular structure. This junctional PACSIN2/EHD4/MICAL-L1 complex controls local VE-cadherin trafficking and coordinates polarized endothelial migration and angiogenesis.\",\n      \"method\": \"Co-immunoprecipitation, fluorescence microscopy, siRNA/shRNA knockdown, VE-cadherin trafficking assays, angiogenesis sprouting assays\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP, localization, functional trafficking and angiogenesis assays, single lab\",\n      \"pmids\": [\"33972531\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"MICAL-L1 depletion impairs delivery of a subset of cargo proteins to the cell surface. The MICAL-L1 RBD domain contributes to PACSIN-mediated membrane tubulation in vitro. Two hydrophobic residues at the MICAL-L1 C-terminus are important for phosphatidic acid binding and for association with membrane tubules. MICAL-L1 associates with Golgi apparatus markers and recycling endosomes.\",\n      \"method\": \"shRNA knockdown, synchronized secretion assay, in vitro membrane tubulation assay with recombinant MICAL-L1-RBD, confocal and STORM super-resolution microscopy, site-directed mutagenesis of C-terminal hydrophobic residues\",\n      \"journal\": \"Biology open\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — in vitro tubulation, mutagenesis, functional secretion assay, single lab with multiple methods\",\n      \"pmids\": [\"34100897\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"MICAL-L1 directly interacts with FCHSD2 and recruits it to the endosomal membrane. FCHSD2 recruitment by MICAL-L1 is required for ARP2/3-mediated branched actin generation, endosome fission, and receptor recycling. Because MICAL-L1 recruits FCHSD2 prior to EHD1, MICAL-L1 orchestrates the sequential steps of endosomal fission by bridging early actin-driven constriction (via FCHSD2/ARP2/3) and subsequent nucleotide hydrolysis/fission (via EHD1).\",\n      \"method\": \"Co-immunoprecipitation, siRNA knockdown of MICAL-L1 and FCHSD2, ARP2/3 actin branching assays, endosome fission assays, receptor recycling assays\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP, knockdown with defined mechanistic phenotypes, actin branching assay, single lab\",\n      \"pmids\": [\"39382837\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"CD2AP and CIN85 are novel constituents of tubular recycling endosomes recruited by MICAL-L1 via their SH3 domains. Depletion of either CD2AP or CIN85 impairs recycling endosome function. MICAL-L1 scaffolding of these proteins likely impacts recycling through effects on the actin cytoskeleton.\",\n      \"method\": \"Co-immunoprecipitation (SH3 domain interactions), siRNA knockdown of CD2AP and CIN85, receptor recycling assays, fluorescence microscopy\",\n      \"journal\": \"Traffic (Copenhagen, Denmark)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — co-IP showing SH3-mediated interaction, knockdown with recycling phenotype, single lab\",\n      \"pmids\": [\"40740057\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"MICAL-L1 is necessary for ETEN (elongated tubular endosomal network) remodeling originating from the endocytic recycling compartment in human dendritic cells, as demonstrated by knockdown experiments.\",\n      \"method\": \"siRNA knockdown, fluorescence microscopy\",\n      \"journal\": \"Communicative & integrative biology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single knockdown experiment, single localization readout, single lab\",\n      \"pmids\": [\"26478765\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"MICAL-L1 functions as a multi-valent endosomal scaffold protein on tubular recycling endosomes (TREs), where its C-terminal coiled-coil domain mediates membrane association via phosphatidic acid binding; it acts as a Rab35 effector that bridges upstream GTPases (Rab35, Arf6) with downstream effectors (Rab8a, Rab13, Rab36), recruits EHD1 and syndapin2 for TRE biogenesis, recruits FCHSD2/ARP2/3 for early actin-driven endosome fission followed by EHD1-mediated nucleotide hydrolysis fission, scaffolds CD2AP, CIN85, and GRAF1 for TRE vesiculation, coordinates Src and VE-cadherin trafficking to regulate cell migration and angiogenesis, and also localizes to centrosomes (anchored by tubulins) to recruit EHD1 for ciliogenesis.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"MICALL1 (MICAL-L1) is a multivalent membrane scaffold that organizes tubular recycling endosomes (TREs) and coordinates receptor recycling, with downstream consequences for cell migration, mitosis, and ciliogenesis [#0, #1]. Its C-terminal coiled-coil region is necessary and sufficient for targeting to tubular membranes, and membrane association is reinforced by direct phosphatidic acid binding through C-terminal hydrophobic residues [#0, #2, #10]. MICAL-L1 acts as a Rab35 effector that bridges upstream GTPases to downstream traffic: Rab35 and Arf6 govern its endosomal recruitment, after which MICAL-L1 scaffolds the non-redundant downstream Rabs Rab8a, Rab13, and Rab36 onto recycling endosomes [#1, #3]. On TREs it nucleates a fission and biogenesis machinery, partnering with the BAR-domain protein syndapin2 to drive membrane tubulation [#2], and orchestrating sequential endosome fission by first recruiting FCHSD2 to trigger ARP2/3-mediated branched actin constriction and then EHD1 for nucleotide-hydrolysis-driven scission [#0, #11]. It additionally scaffolds GRAF1 and the SH3-domain adaptors CD2AP and CIN85 to promote tubule vesiculation and recycling [#4, #12]. Through this trafficking function MICAL-L1 supports growth-factor- and integrin-induced Src activation, focal adhesion turnover, and migration [#6], and within endothelial junctions a PACSIN2/EHD4/MICAL-L1 complex directs VE-cadherin trafficking during angiogenesis [#9]. Beyond recycling, MICAL-L1 binds α/β- and γ-tubulin to anchor at centrosomes where it recruits EHD1 to drive CP110 removal and ciliogenesis [#8], and is required for recycling-endosome-dependent cytokinesis and chromosome alignment in mitosis [#5]. MICALL1 is a transcriptional target of p53 that links DNA damage signaling to TRE biogenesis [#7].\",\n  \"teleology\": [\n    {\n      \"year\": 2009,\n      \"claim\": \"Established MICAL-L1 as the scaffold that physically links EHD1 and Rab8a on tubular recycling endosomes, answering how these recycling factors are co-recruited to tubular membranes.\",\n      \"evidence\": \"Reciprocal co-IP, domain truncation mapping the coiled-coil, siRNA knockdown, and transferrin/integrin recycling assays across cell lines\",\n      \"pmids\": [\"19864458\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not define upstream GTPase control of MICAL-L1 recruitment\", \"No structural detail of the coiled-coil membrane interaction\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Placed MICAL-L1 within a GTPase hierarchy, showing Rab35 and Arf6 act upstream to control its endosomal recruitment and that MICAL-L1/Arf6 lie upstream of Rab8a.\",\n      \"evidence\": \"Co-IP, siRNA knockdown, and active/inactive GTPase mutant overexpression with fluorescence microscopy\",\n      \"pmids\": [\"21951725\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism by which active Rab35 displaces MICAL-L1 from TREs not resolved\", \"Single lab\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Defined the lipid basis of TRE membrane association, showing MICAL-L1 and syndapin2 bind phosphatidic acid and cooperate to nucleate tubulation.\",\n      \"evidence\": \"Lipid-binding assays, in vitro liposome tubulation with recombinant syndapin2, and co-IP\",\n      \"pmids\": [\"23596323\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo contribution of phosphatidic acid to TRE biogenesis not quantified\", \"Did not map the MICAL-L1 lipid-binding residues (later addressed in #10)\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Showed MICAL-L1 scaffolds multiple non-redundant downstream Rabs (Rab8, Rab13, Rab36) during NGF-induced neurite outgrowth, with Rab36/JIP4 having a distinct functional output.\",\n      \"evidence\": \"siRNA knockdown of Rab35/MICAL-L1/Rab8/Rab13/Rab36 and neurite outgrowth assays in PC12 cells\",\n      \"pmids\": [\"25086062\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct binding of each Rab to MICAL-L1 not biochemically dissected\", \"Generality beyond NGF/PC12 system unknown\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Identified GRAF1 as a MICAL-L1/EHD1-associated factor driving TRE vesiculation, linking the scaffold to membrane scission.\",\n      \"evidence\": \"Co-IP, knockdown, and a semi-permeabilized cell vesiculation assay with purified EHD1 and GRAF1\",\n      \"pmids\": [\"25364729\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Order of GRAF1 versus other fission factors not established\", \"Single lab\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Connected MICAL-L1 trafficking function to Src activation, focal adhesion turnover, and migration, defining a physiological output of TRE recycling.\",\n      \"evidence\": \"siRNA knockdown of MICAL-L1/EHD1, Src activation, focal adhesion turnover, and migration assays in HeLa and fibroblasts\",\n      \"pmids\": [\"24481818\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct cargo-binding mechanism for Src vesicles not shown\", \"Single lab\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Revealed mitotic roles, including an EHD1-independent function in chromosome alignment and an EHD1-dependent recycling role in cytokinesis.\",\n      \"evidence\": \"Differential siRNA knockdown of MICAL-L1 and EHD1 with binucleation, chromosome alignment, and microtubule dynamics readouts\",\n      \"pmids\": [\"25287187\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular partner for the EHD1-independent mitotic function unidentified\", \"Mechanism linking MICAL-L1 to microtubule dynamics unclear\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Identified MICALL1 as a direct p53 transcriptional target, linking DNA damage signaling to TRE biogenesis.\",\n      \"evidence\": \"Multi-omics, p53-binding motif identification ~3 kb upstream, siRNA depletion of p53/MICALL1, and co-localization with RAB8A/CD2AP\",\n      \"pmids\": [\"28714518\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional consequence of p53-driven TRE biogenesis for DNA damage outcomes not established\", \"Single lab\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Extended MICAL-L1 function to the centrosome/cilium, showing it binds tubulins to anchor at centrioles and recruit EHD1 for CP110 removal and ciliogenesis.\",\n      \"evidence\": \"siRNA knockdown with ciliogenesis phenotypes, mass spectrometry, and direct binding assays to α/β- and γ-tubulin\",\n      \"pmids\": [\"31615969\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"How a recycling scaffold also acts at centrioles mechanistically distinct from TREs unclear\", \"Single lab\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Demonstrated a junctional PACSIN2/EHD4/MICAL-L1 complex controlling VE-cadherin trafficking, polarized endothelial migration, and angiogenesis.\",\n      \"evidence\": \"Co-IP, knockdown, VE-cadherin trafficking assays, and angiogenesis sprouting assays in endothelial cells\",\n      \"pmids\": [\"33972531\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether this is the same TRE machinery repurposed at junctions not fully resolved\", \"Single lab\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Mapped C-terminal hydrophobic residues required for phosphatidic acid binding and defined the RBD contribution to PACSIN-mediated tubulation and surface cargo delivery.\",\n      \"evidence\": \"shRNA knockdown, synchronized secretion assay, in vitro tubulation with recombinant MICAL-L1-RBD, STORM imaging, and site-directed mutagenesis\",\n      \"pmids\": [\"34100897\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Identity of the affected cargo subset incompletely defined\", \"Golgi association mechanism not detailed\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Resolved the sequential logic of endosome fission, showing MICAL-L1 recruits FCHSD2/ARP2/3 for early actin-driven constriction before EHD1-mediated scission.\",\n      \"evidence\": \"Co-IP, siRNA knockdown of MICAL-L1/FCHSD2, ARP2/3 actin branching, endosome fission, and recycling assays\",\n      \"pmids\": [\"39382837\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Temporal coordination measured indirectly\", \"Single lab\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Added CD2AP and CIN85 as SH3-domain-recruited TRE constituents acting through the actin cytoskeleton in recycling.\",\n      \"evidence\": \"Co-IP mapping SH3 interactions, siRNA knockdown of CD2AP/CIN85, and receptor recycling assays\",\n      \"pmids\": [\"40740057\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct actin-cytoskeleton link inferred, not demonstrated\", \"Single lab\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How MICAL-L1's many partner interactions are spatially and temporally segregated across TREs, centrosomes, junctions, and the mitotic apparatus remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural model of the multivalent scaffold with bound partners\", \"Regulation of context-specific complex assembly unknown\", \"Whether disease-relevant phenotypes arise from MICALL1 loss not established\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 1, 3, 4, 11, 12]},\n      {\"term_id\": \"GO:0008289\", \"supporting_discovery_ids\": [2, 10]},\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [8]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005768\", \"supporting_discovery_ids\": [0, 1, 2, 4, 11, 12]},\n      {\"term_id\": \"GO:0005815\", \"supporting_discovery_ids\": [8]},\n      {\"term_id\": \"GO:0005929\", \"supporting_discovery_ids\": [8]},\n      {\"term_id\": \"GO:0005794\", \"supporting_discovery_ids\": [10]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-5653656\", \"supporting_discovery_ids\": [0, 4, 11, 12]},\n      {\"term_id\": \"R-HSA-9609507\", \"supporting_discovery_ids\": [6, 9, 10]},\n      {\"term_id\": \"R-HSA-1852241\", \"supporting_discovery_ids\": [8]}\n    ],\n    \"complexes\": [\n      \"MICAL-L1/EHD1/Rab8a TRE scaffold\",\n      \"PACSIN2/EHD4/MICAL-L1 junctional complex\",\n      \"MICAL-L1/EHD1/GRAF1 complex\"\n    ],\n    \"partners\": [\n      \"EHD1\",\n      \"RAB8A\",\n      \"RAB35\",\n      \"ARF6\",\n      \"PACSIN2\",\n      \"FCHSD2\",\n      \"CD2AP\",\n      \"CIN85\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":8,"faith_total":8,"faith_pct":100.0}}