{"gene":"MICALL2","run_date":"2026-06-10T02:59:50","timeline":{"discoveries":[{"year":2006,"finding":"MICAL-L2 (JRAB) was identified as a Rab13 effector protein that specifically binds the GTP-bound form of Rab13 via its C-terminal coiled-coil domain, localizes at tight junctions in epithelial cells, and mediates endocytic recycling of occludin (but not transferrin receptor). A MICAL-L2 mutant lacking the Rab13-binding domain (MICAL-L2-N) specifically inhibited occludin recycling and blocked TJ formation in Ca2+-switch assays. MICAL-L2 was displaced from TJs upon actin depolymerization and redistributed along actin structures, indicating it links Rab13 to the actin cytoskeleton.","method":"Immunoprecipitation, immunofluorescence, recycling assay, Ca2+-switch assay, yeast two-hybrid (initial identification), dominant-negative mutant expression","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, multiple orthogonal functional assays (recycling, Ca2+-switch, dominant-negative), replicated in subsequent studies","pmids":["16525024"],"is_preprint":false},{"year":2007,"finding":"JRAB/MICAL-L2 interacts with both Rab8 and Rab13 via its C-terminal domain, and these two Rab GTPases compete with each other for MICAL-L2 binding. Rab8 and Rab13 functionally associate with MICAL-L2 at perinuclear recycling/storage compartments and at the plasma membrane, respectively. Rab13 knockdown suppressed claudin-1 and occludin but not E-cadherin transport, while Rab8 knockdown inhibited Rab13-independent E-cadherin transport; MICAL-L2 knockdown or dominant-negative MICAL-L2-C expression inhibited transport of all three junctional proteins, coordinating TJ and AJ assembly.","method":"siRNA knockdown, dominant-negative mutant expression, Ca2+-switch assay, immunofluorescence, co-immunoprecipitation","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (KD, dominant-negative, Co-IP, imaging), builds on prior replicated findings","pmids":["18094055"],"is_preprint":false},{"year":2008,"finding":"Actinin-4 was identified as a binding partner for the plasma membrane-targeting domain of JRAB/MICAL-L2 using yeast two-hybrid and confirmed by co-immunoprecipitation. Actinin-4 colocalizes with JRAB/MICAL-L2 at cell-cell junctions and links JRAB/MICAL-L2 to F-actin. The actinin-4–MICAL-L2 interaction was enhanced by Rab13 activation. siRNA depletion of actinin-4 delayed JRAB/MICAL-L2 recruitment to cell-cell junctions and impaired functional TJ formation during epithelial polarization.","method":"Yeast two-hybrid, co-immunoprecipitation, siRNA knockdown, Ca2+-switch assay, immunofluorescence","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — yeast two-hybrid plus reciprocal Co-IP plus siRNA functional validation, multiple orthogonal methods","pmids":["18332111"],"is_preprint":false},{"year":2008,"finding":"JRAB/MICAL-L2 domain structure was characterized: it contains a calponin-homology (CH) domain at the N-terminus, a LIM domain in the middle, and a coiled-coil domain at the C-terminus. The C-terminus specifically binds the GTP-bound form of Rab13. In epithelial cells it localizes to tight junctions; in fibroblasts it distributes along stress fibers.","method":"Domain deletion analysis, immunoprecipitation, immunofluorescence, yeast two-hybrid (referenced prior work)","journal":"Methods in enzymology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — consolidates prior structural characterization data, single review/methods paper but based on earlier experimental work","pmids":["18413246"],"is_preprint":false},{"year":2009,"finding":"JRAB/MICAL-L2 exhibits an intramolecular interaction between its N-terminal CH and LIM domains and the C-terminal coiled-coil domain. Rab13 binding to JRAB/MICAL-L2 stimulates interaction of MICAL-L2 with actinin-4, which localizes to cell body and neurite tips in PC12 cells. Expression of MICAL-L2 alone inhibits neurite outgrowth, but co-expression of dominant-active Rab13 rescues this phenotype, suggesting Rab13-dependent conformational change enables transfer of actinin-4 to neurite tips to reorganize the actin cytoskeleton.","method":"Overexpression/dominant-active mutants, co-immunoprecipitation, immunofluorescence, neurite outgrowth assay in PC12 cells","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 2 / Moderate — intramolecular interaction mapped by deletion/co-IP, functional rescue with dominant-active Rab13, multiple orthogonal methods in single study","pmids":["20008558"],"is_preprint":false},{"year":2007,"finding":"Rab13 and JRAB/MICAL-L2 are involved in epithelial cell scattering in response to TPA. During TPA-induced MDCK cell scattering, Rab13 was transiently activated, and both Rab13 and JRAB/MICAL-L2 co-localized with F-actin at cell-cell contact sites before accumulating at lamellipodia. Knockdown of JRAB/MICAL-L2 inhibited TPA-induced scattering, rescued by re-expression of mouse JRAB/MICAL-L2.","method":"siRNA knockdown, rescue by re-expression, immunofluorescence, Rab13 activation assay","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — siRNA KD with rescue experiment, colocalization, single lab","pmids":["17891173"],"is_preprint":false},{"year":2012,"finding":"JRAB/MICAL-L2 interacts with both actinin-1 and actinin-4 and with filamentous actin (F-actin) via different domains, and regulates actin cross-linking and stabilization. During epithelial junctional development, JRAB is enriched in the actin bundle at the free border and undergoes a Rab13-dependent conformational change required for maturation of cell-cell adhesion.","method":"Co-immunoprecipitation, domain deletion analysis, immunofluorescence, live imaging, Rab13 dominant-active/dominant-negative mutants","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple binding partners mapped with Co-IP/domain analysis, Rab13-dependent conformational change demonstrated, consistent with prior replicated findings","pmids":["23100251"],"is_preprint":false},{"year":2013,"finding":"In the 'open' conformation (JRABΔCC mutant), JRAB/MICAL-L2 co-immunoprecipitates with filamin (an actin cross-linking protein). Expression of ASB2, which induces degradation of all three filamin isoforms, inhibited JRABΔCC-induced cell spreading with membrane ruffles, demonstrating that JRAB regulates actin cytoskeletal reorganization and cell spreading through filamins. In contrast, actinin-1/-4 co-precipitated with JRABΔCC but did not affect cell spreading.","method":"Co-immunoprecipitation with conformational mutants, ASB2-mediated filamin degradation, morphological analysis in NIH3T3 cells","journal":"Genes to cells","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP with functional validation via filamin depletion, single lab, two methods","pmids":["23890175"],"is_preprint":false},{"year":2015,"finding":"Rab13 engages MICAL-L2 as a scaffold upon insulin stimulation in muscle cells. Insulin increased Rab13 binding to MICAL-L2 and binding of MICAL-L2 to α-actinin-4 (ACTN4), forming an insulin-dependent ternary complex (Rab13–MICAL-L2–ACTN4). GLUT4 associated with this complex in response to insulin via the ACTN4-binding domain of MICAL-L2. Knockdown of MICAL-L2 or expression of truncated MICAL-L2-CT impaired insulin-stimulated GLUT4 translocation to the plasma membrane.","method":"Co-immunoprecipitation, pull-down with MICAL-L2 fragments, siRNA knockdown, confocal fluorescence, TIRF microscopy, structured illumination microscopy","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (pull-down, Co-IP, TIRF/SIM imaging, siRNA KD), domain mapping of ACTN4 interaction, consistent functional readout","pmids":["26538022"],"is_preprint":false},{"year":2016,"finding":"JRAB/MICAL-L2 undergoes a Rab13-dependent conformational change between closed and open states. A structural model was generated by bioinformatic and biochemical analyses. Impairment of MICAL-L2 conformational plasticity (using fixed conformation mutants) caused excessive rigidity and loss of directionality in collective cell migration. Live imaging and computational analysis showed that closed/open conformations dictate distinct collective migration behaviors.","method":"Structural modeling, biochemical conformation assays, live imaging, computational biomechanics, mutant expression","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 2 / Moderate — structural model validated biochemically, functional consequence of conformational change demonstrated by live imaging and quantitative cell biology; multiple orthogonal methods","pmids":["27582384"],"is_preprint":false},{"year":2019,"finding":"The first LIM zinc finger domain of JRAB/MICAL-L2 binds the first and second actin molecules at the minus end of F-actin, potentially inhibiting actin filament depolymerization. This same zinc finger domain also contributes to the intramolecular interaction with the C-terminal coiled-coil domain, and residues responsible for intramolecular interaction overlap with those involved in F-actin association, providing a mechanism for fine-tuning JRAB function.","method":"Biochemical actin-binding assays, mutagenesis, intramolecular interaction mapping, co-immunoprecipitation with mutants","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 1-2 / Moderate — actin-binding assay with mutagenesis, intramolecular interaction mapped, single lab","pmids":["31488862"],"is_preprint":false},{"year":2019,"finding":"MICAL-L2 expression upregulated EGFR protein level through a transcription-independent mechanism involving inhibition of EGFR degradation in lysosomes. This effect on EGFR stability was mediated in a Cdc42-dependent manner, activating downstream HSP27/cytoskeleton and HSP27/β-catenin signaling pathways to promote gastric cancer cell migration.","method":"siRNA knockdown, overexpression, Western blotting, lysosome inhibitor experiments, Cdc42 activation assays, immunofluorescence","journal":"Journal of cellular and molecular medicine","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — single lab, multiple assays but mechanistic basis of Cdc42 dependence not fully reconstituted in vitro","pmids":["31034158"],"is_preprint":false},{"year":2021,"finding":"MICAL-L2 physically interacts with c-Myc protein (co-immunoprecipitation), maintains nuclear c-Myc levels, prolongs its half-life, and inhibits c-Myc polyubiquitination. MICAL-L2 knockdown accelerated c-Myc degradation through polyubiquitylation; MICAL-L2 blocked c-Myc phosphorylation at Thr58, which is required for ubiquitin-dependent c-Myc degradation, thereby acting as a deubiquitination/stabilization factor for c-Myc in NSCLC cells.","method":"Co-immunoprecipitation, immunofluorescence, polyubiquitylation detection assay, protein stability assay, siRNA knockdown, Western blotting","journal":"Frontiers in cell and developmental biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP confirmed interaction, ubiquitination assays and Thr58 phosphorylation mechanistically linked, single lab","pmids":["33520979"],"is_preprint":false},{"year":2022,"finding":"TRIM21 (ubiquitin E3 ligase) mediates ubiquitination and proteasome-dependent degradation of MICALL2, acting as a negative regulator. Co-immunoprecipitation and mass spectrometry identified TRIM21 as a MICALL2-interacting protein. TRIM21 expression negatively correlates with MICALL2 levels and reversely regulates MICALL2-dependent tumorigenic activity (growth and migration via Wnt/β-catenin) in colorectal cancer cells.","method":"Co-immunoprecipitation, mass spectrometry, ubiquitination assays, siRNA/overexpression functional assays, Western blotting","journal":"Cell communication and signaling","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP plus mass spectrometry identification, ubiquitination assay confirms TRIM21 as E3 ligase, single lab","pmids":["36307841"],"is_preprint":false},{"year":2023,"finding":"MICALL2 silencing in ovarian cancer cells promoted EGFR lysosomal degradation in a Rac1-dependent (rather than Cdc42-dependent as in gastric cancer) manner. Rac1 suppression attenuated MICALL2-induced pro-EGFR, pro-MMP9, and pro-invasion effects. MMP9 was identified as the target gene downstream of MICALL2 via EGFR-AKT-mTOR signaling, regulating invadopodium formation and cell invasion.","method":"siRNA knockdown, overexpression, Rac1 inhibition, autophagy inhibitors (acadesine, chloroquine), co-immunoprecipitation, invasion assays","journal":"International journal of molecular sciences","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — mechanistic pathway defined by pharmacological inhibition and KD, single lab, no in vitro reconstitution","pmids":["38203692"],"is_preprint":false},{"year":2024,"finding":"MICAL-L2 stabilizes ACTN4 protein in a Rab13-dependent manner in clear cell renal cell carcinoma (KIRC) cells, reducing ACTN4 degradation. This MICAL-L2–ACTN4 complex interaction was confirmed by co-immunoprecipitation. ACTN4 stabilization leads to increased Vimentin expression, promoting cancer progression and resistance to sunitinib and everolimus.","method":"Co-immunoprecipitation, siRNA knockdown, overexpression, Western blotting, wound-healing, Transwell, CCK-8 assays","journal":"Biochimica et biophysica acta. Molecular basis of disease","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — Co-IP confirmed MICAL-L2/ACTN4 interaction with Rab13 dependence, functional assays, single lab","pmids":["39689763"],"is_preprint":false},{"year":2025,"finding":"MICALL2 prevents β-catenin from binding to GSK3β, thereby shielding β-catenin from degradation and promoting Wnt/β-catenin signaling in hepatic stellate cells. A small molecule IMB16-5 binds to the C-terminal domain of MICALL2 (identified by LiP-SMap and molecular dynamics simulation) and disrupts the MICALL2–β-catenin interaction, suppressing β-catenin nuclear translocation and HSC activation. MICALL2 overexpression in vivo abolished the therapeutic efficacy of IMB16-5, confirming target engagement.","method":"LiP-SMap (limited proteolysis-small molecule mapping), molecular dynamics simulation, co-immunoprecipitation, in vivo mouse fibrosis model, overexpression rescue experiment","journal":"International immunopharmacology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — LiP-SMap maps drug binding site, Co-IP shows MICALL2-β-catenin-GSK3β relationship, in vivo confirmation, single lab","pmids":["41461119"],"is_preprint":false},{"year":2026,"finding":"PSMD14 (a proteasome deubiquitinase) physically interacts with MICALL2 (confirmed by co-immunoprecipitation) and suppresses MICALL2 ubiquitination and degradation, thereby increasing MICALL2 protein levels. DUSP4-mediated dephosphorylation of PSMD14 enhances PSMD14–MICALL2 interaction and amplifies PSMD14's stabilizing effect on MICALL2, promoting ccRCC malignant progression.","method":"Co-immunoprecipitation, immunoprecipitation for ubiquitination analysis, PSMD14 phosphorylation analysis, siRNA knockdown, Western blotting, in vivo xenograft model","journal":"Journal of translational medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP confirmed PSMD14–MICALL2 interaction, ubiquitination and phosphorylation assays, functional validation in vivo, single lab","pmids":["42174657"],"is_preprint":false},{"year":2025,"finding":"MICALL2 was identified as a candidate substrate of LRRK2 kinase in proximity-labeling mass spectrometry and quantitative phosphoproteomics experiments. LRRK2 inhibitors altered MICALL2 phosphorylation status at the proteome-wide level.","method":"Proximity labeling mass spectrometry (BioID/TurboID), quantitative phosphoproteomics, LRRK1/2 inhibitor treatment","journal":"bioRxiv","confidence":"Low","confidence_rationale":"Tier 4 / Weak — identified as candidate substrate in large-scale screen, no direct in vitro kinase assay or site-specific mutagenesis confirming LRRK2-mediated phosphorylation of MICALL2","pmids":["bio_10.1101_2025.09.03.674114"],"is_preprint":true}],"current_model":"MICALL2/JRAB is a multidomain scaffold protein (CH–LIM–coiled-coil) that functions as an effector of GTP-bound Rab13 (and Rab8) to coordinate vesicle trafficking and actin cytoskeletal reorganization: in the Rab13-bound open conformation it recruits actinin-1/4 and filamins to regulate actin cross-linking and stabilization, mediates endocytic recycling of tight junction proteins (occludin, claudins) and E-cadherin to the plasma membrane, undergoes a Rab13-dependent intramolecular conformational switch that controls collective cell migration directionality, and also acts in non-junctional contexts to stabilize EGFR (via Cdc42/Rac1-dependent inhibition of lysosomal degradation), stabilize c-Myc (by blocking Thr58 phosphorylation and polyubiquitination), protect β-catenin from GSK3β-mediated degradation to activate Wnt signaling, and stabilize ACTN4 in a Rab13-dependent manner; its own protein stability is regulated by TRIM21-mediated ubiquitination/proteasomal degradation and by PSMD14-mediated deubiquitination."},"narrative":{"mechanistic_narrative":"MICALL2 (JRAB) is a multidomain scaffold protein (CH–LIM–coiled-coil) that acts as an effector of GTP-bound Rab13, and also Rab8, to couple vesicle trafficking with actin cytoskeletal reorganization at epithelial junctions [PMID:16525024, PMID:18094055, PMID:18413246]. Through its C-terminal coiled-coil it binds active Rab13/Rab8, and through its membrane-targeting and LIM regions it engages the actin cross-linkers actinin-1/4 and filamins and binds F-actin directly, thereby regulating actin cross-linking and stabilization [PMID:18332111, PMID:23100251, PMID:23890175, PMID:31488862]. In this capacity it mediates endocytic recycling of tight- and adherens-junction proteins—occludin, claudin-1, and E-cadherin—to the plasma membrane and is required for coordinated TJ/AJ assembly [PMID:16525024, PMID:18094055]. MICALL2 operates as a conformational switch: an intramolecular interaction between its N-terminal CH/LIM domains and the C-terminal coiled-coil holds it closed, and Rab13 binding opens the molecule to transfer actinin-4 onto actin structures; this Rab13-dependent open/closed plasticity governs actin remodeling, junctional maturation, and the directionality of collective cell migration [PMID:20008558, PMID:23100251, PMID:27582384, PMID:31488862]. Beyond junctions, MICALL2 serves as a Rab13-dependent scaffold linking GLUT4 to a Rab13–MICALL2–ACTN4 ternary complex during insulin-stimulated GLUT4 translocation [PMID:26538022]. In cancer contexts MICALL2 acts as a stability factor for multiple oncogenic effectors—stabilizing EGFR by blocking its lysosomal degradation via Cdc42- or Rac1-dependent signaling [PMID:31034158, PMID:38203692], stabilizing c-Myc by preventing Thr58 phosphorylation and polyubiquitination [PMID:33520979], shielding β-catenin from GSK3β to activate Wnt signaling [PMID:41461119], and stabilizing ACTN4 in a Rab13-dependent manner [PMID:39689763]. MICALL2's own abundance is set by opposing ubiquitin machinery: TRIM21 ubiquitinates it for proteasomal degradation, while the proteasome-associated deubiquitinase PSMD14 stabilizes it [PMID:36307841, PMID:42174657].","teleology":[{"year":2006,"claim":"Established MICALL2 as a Rab13 effector that links a small GTPase to junctional vesicle trafficking, answering how Rab13 controls tight junction assembly.","evidence":"Co-IP, recycling and Ca2+-switch assays with a Rab13-binding-deficient mutant in epithelial cells","pmids":["16525024"],"confidence":"High","gaps":["Did not resolve the structural basis of GTP-dependent Rab13 binding","Cargo selectivity (occludin vs transferrin receptor) mechanism unexplained"]},{"year":2007,"claim":"Showed MICALL2 integrates two Rab GTPases by competitive binding, explaining how it coordinates both tight- and adherens-junction protein delivery.","evidence":"siRNA knockdown, dominant-negative MICAL-L2-C, Ca2+-switch and Co-IP in epithelial cells","pmids":["18094055"],"confidence":"High","gaps":["Did not define how Rab8 vs Rab13 compartment selection is regulated","Spatial control of competition not addressed"]},{"year":2007,"claim":"Linked MICALL2/Rab13 to epithelial scattering, extending its role from static junctions to migratory behavior.","evidence":"siRNA knockdown with re-expression rescue and Rab13 activation assay in TPA-stimulated MDCK cells","pmids":["17891173"],"confidence":"Medium","gaps":["Did not identify the molecular trigger coupling Rab13 activation to scattering","Single-lab observation"]},{"year":2008,"claim":"Identified actinin-4 as the link between MICALL2 and F-actin and defined the CH–LIM–coiled-coil domain architecture, providing the structural framework for its scaffold function.","evidence":"Yeast two-hybrid, Co-IP, siRNA, domain-deletion analysis and immunofluorescence","pmids":["18332111","18413246"],"confidence":"High","gaps":["Precise actinin-4-binding domain boundaries partially defined","How Rab13 enhances the actinin-4 interaction left mechanistically open"]},{"year":2009,"claim":"Discovered the Rab13-driven intramolecular conformational switch, explaining how Rab13 binding mobilizes actinin-4 to remodel actin.","evidence":"Co-IP, dominant-active Rab13, neurite-outgrowth rescue in PC12 cells","pmids":["20008558"],"confidence":"High","gaps":["Atomic structure of open vs closed states not determined","Kinetics of the conformational transition unknown"]},{"year":2012,"claim":"Demonstrated MICALL2 binds actinin-1/4 and F-actin through distinct domains to control actin cross-linking and junctional maturation, consolidating its cytoskeletal-regulatory role.","evidence":"Co-IP, domain deletion, live imaging with Rab13 mutants","pmids":["23100251"],"confidence":"High","gaps":["Quantitative effect on actin cross-linking not measured in vitro","Hierarchy of multiple actin-binding partners unresolved"]},{"year":2013,"claim":"Showed the open conformation recruits filamins to drive cell spreading, distinguishing functional outputs of different MICALL2-bound cross-linkers.","evidence":"Co-IP with conformational mutants and ASB2-mediated filamin degradation in NIH3T3 cells","pmids":["23890175"],"confidence":"Medium","gaps":["Why actinins co-precipitate but do not affect spreading unexplained","Single-lab, two-method support"]},{"year":2015,"claim":"Extended the scaffold function to metabolic trafficking by placing MICALL2 in an insulin-induced Rab13–MICALL2–ACTN4 complex that delivers GLUT4.","evidence":"Pull-down, Co-IP, siRNA, TIRF/SIM imaging in muscle cells","pmids":["26538022"],"confidence":"High","gaps":["How insulin signaling triggers Rab13–MICALL2 binding not defined","Direct GLUT4 contact vs indirect not resolved"]},{"year":2016,"claim":"Built a structural model of the conformational switch and tied conformational plasticity directly to collective migration directionality.","evidence":"Structural modeling, biochemical conformation assays, live imaging and computational biomechanics with fixed-conformation mutants","pmids":["27582384"],"confidence":"High","gaps":["Model not validated by experimental high-resolution structure","Upstream control of conformational state during migration unclear"]},{"year":2019,"claim":"Defined a direct F-actin-binding mechanism for the LIM zinc finger and showed overlap with the intramolecular interface, providing a molecular basis for fine-tuning MICALL2 activity.","evidence":"Biochemical actin-binding assays, mutagenesis, intramolecular interaction mapping","pmids":["31488862"],"confidence":"Medium","gaps":["Functional consequence of minus-end binding on filament dynamics not tested in cells","Single-lab biochemistry"]},{"year":2019,"claim":"Opened a non-junctional oncogenic role by showing MICALL2 stabilizes EGFR via Cdc42-dependent inhibition of lysosomal degradation.","evidence":"Knockdown/overexpression, lysosome inhibitors, Cdc42 activation assays in gastric cancer cells","pmids":["31034158"],"confidence":"Medium","gaps":["Direct biochemical link between MICALL2 and EGFR trafficking not reconstituted","Cdc42-dependence mechanism incomplete"]},{"year":2021,"claim":"Identified MICALL2 as a c-Myc stabilizer that blocks Thr58 phosphorylation and polyubiquitination, broadening its function to nuclear oncoprotein regulation.","evidence":"Co-IP, polyubiquitylation and protein-stability assays, siRNA in NSCLC cells","pmids":["33520979"],"confidence":"Medium","gaps":["Whether the interaction is direct or how it blocks Thr58 phosphorylation unresolved","Single-lab"]},{"year":2022,"claim":"Defined upstream control of MICALL2 abundance by identifying TRIM21 as its degradative E3 ligase.","evidence":"Co-IP, mass spectrometry, ubiquitination assays and functional readouts in colorectal cancer cells","pmids":["36307841"],"confidence":"Medium","gaps":["TRIM21 ubiquitination site(s) on MICALL2 not mapped","Single-lab"]},{"year":2023,"claim":"Showed EGFR stabilization can proceed through Rac1 rather than Cdc42, with MMP9 as a downstream effector, indicating context-dependent GTPase usage.","evidence":"Knockdown/overexpression, Rac1 inhibition, autophagy inhibitors, invasion assays in ovarian cancer cells","pmids":["38203692"],"confidence":"Medium","gaps":["Basis for tissue-specific Cdc42 vs Rac1 choice unexplained","No in vitro reconstitution"]},{"year":2024,"claim":"Demonstrated Rab13-dependent stabilization of ACTN4 links MICALL2 to vimentin upregulation and therapy resistance, connecting its scaffold role to malignant progression.","evidence":"Co-IP, siRNA/overexpression and functional assays in clear cell renal cell carcinoma cells","pmids":["39689763"],"confidence":"Medium","gaps":["Mechanism by which MICALL2 protects ACTN4 from degradation undefined","Single-lab"]},{"year":2025,"claim":"Established that MICALL2 shields β-catenin from GSK3β to activate Wnt signaling and identified a small molecule binding its C-terminus that disrupts this function.","evidence":"LiP-SMap, molecular dynamics, Co-IP and in vivo fibrosis model with overexpression rescue","pmids":["41461119"],"confidence":"Medium","gaps":["Direct vs scaffolded MICALL2–β-catenin contact not fully resolved","Single-lab"]},{"year":2026,"claim":"Completed the regulatory loop on MICALL2 stability by showing the deubiquitinase PSMD14, controlled by DUSP4-mediated dephosphorylation, stabilizes MICALL2.","evidence":"Co-IP, ubiquitination and phosphorylation assays, siRNA, in vivo xenograft in ccRCC","pmids":["42174657"],"confidence":"Medium","gaps":["Whether TRIM21 and PSMD14 act on the same ubiquitin sites unknown","Single-lab"]},{"year":2025,"claim":"Flagged MICALL2 as a candidate LRRK2 kinase substrate, raising the possibility of phosphoregulation of the scaffold.","evidence":"Proximity-labeling MS and quantitative phosphoproteomics with LRRK inhibitors (preprint)","pmids":["bio_10.1101_2025.09.03.674114"],"confidence":"Low","gaps":["No direct in vitro kinase assay or site-specific mutagenesis confirming LRRK2-mediated phosphorylation","Functional consequence of phosphorylation unknown"]},{"year":null,"claim":"How the Rab13-driven conformational switch is mechanistically coupled to MICALL2's diverse protein-stabilization activities (EGFR, c-Myc, β-catenin, ACTN4) remains unresolved.","evidence":"","pmids":[],"confidence":"Low","gaps":["No high-resolution structure of full-length MICALL2 in either conformation","Whether stabilization activities are direct or scaffold-dependent unestablished","Upstream signals selecting junctional vs oncogenic functions undefined"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,2,6,8]},{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[2,6,7,10]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[11,12,15,16]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[0,1,8]},{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[0,3,6]},{"term_id":"GO:0031410","term_label":"cytoplasmic vesicle","supporting_discovery_ids":[0,1]}],"pathway":[{"term_id":"R-HSA-5653656","term_label":"Vesicle-mediated transport","supporting_discovery_ids":[0,1,8]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[11,16]},{"term_id":"R-HSA-1500931","term_label":"Cell-Cell communication","supporting_discovery_ids":[0,1,2]}],"complexes":["Rab13–MICALL2–ACTN4 ternary complex"],"partners":["RAB13","RAB8A","ACTN4","ACTN1","TRIM21","PSMD14","MYC","CTNNB1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q8IY33","full_name":"MICAL-like protein 2","aliases":["Junctional Rab13-binding protein","Molecule interacting with CasL-like 2","MICAL-L2"],"length_aa":904,"mass_kda":97.5,"function":"Effector of small Rab GTPases which is involved in junctional complexes assembly through the regulation of cell adhesion molecules transport to the plasma membrane and actin cytoskeleton reorganization. Regulates the endocytic recycling of occludins, claudins and E-cadherin to the plasma membrane and may thereby regulate the establishment of tight junctions and adherens junctions. In parallel, may regulate actin cytoskeleton reorganization directly through interaction with F-actin or indirectly through actinins and filamins. Most probably involved in the processes of epithelial cell differentiation, cell spreading and neurite outgrowth (By similarity). Undergoes liquid-liquid phase separation to form tubular recycling endosomes. Plays 2 sequential roles in the biogenesis of tubular recycling endosomes: first organizes phase separation and then the closed form formed by interaction with RAB8A promotes endosomal tubulation (By similarity)","subcellular_location":"Cell membrane; Cell junction, tight junction; Recycling endosome; Cell projection; Cytoplasm, cytoskeleton","url":"https://www.uniprot.org/uniprotkb/Q8IY33/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/MICALL2","classification":"Not Classified","n_dependent_lines":2,"n_total_lines":1208,"dependency_fraction":0.0016556291390728477},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/MICALL2","total_profiled":1310},"omim":[{"mim_id":"620912","title":"MICAL-LIKE PROTEIN 2; MICALL2","url":"https://www.omim.org/entry/620912"},{"mim_id":"602672","title":"RAS-ASSOCIATED PROTEIN RAB13; RAB13","url":"https://www.omim.org/entry/602672"},{"mim_id":"165040","title":"RAS-ASSOCIATED PROTEIN RAB8A; RAB8A","url":"https://www.omim.org/entry/165040"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in many","driving_tissues":[],"url":"https://www.proteinatlas.org/search/MICALL2"},"hgnc":{"alias_symbol":["MGC46023","FLJ23471","MICAL-L2","JRAB"],"prev_symbol":[]},"alphafold":{"accession":"Q8IY33","domains":[{"cath_id":"1.10.418.10","chopping":"2-108","consensus_level":"high","plddt":88.9661,"start":2,"end":108},{"cath_id":"2.10.110.10","chopping":"184-239","consensus_level":"medium","plddt":82.0014,"start":184,"end":239},{"cath_id":"-","chopping":"735-889","consensus_level":"medium","plddt":85.3225,"start":735,"end":889}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8IY33","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q8IY33-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q8IY33-F1-predicted_aligned_error_v6.png","plddt_mean":55.31},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=MICALL2","jax_strain_url":"https://www.jax.org/strain/search?query=MICALL2"},"sequence":{"accession":"Q8IY33","fasta_url":"https://rest.uniprot.org/uniprotkb/Q8IY33.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q8IY33/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8IY33"}},"corpus_meta":[{"pmid":"18094055","id":"PMC_18094055","title":"The interaction of JRAB/MICAL-L2 with Rab8 and Rab13 coordinates the assembly of tight junctions and adherens junctions.","date":"2007","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/18094055","citation_count":102,"is_preprint":false},{"pmid":"16525024","id":"PMC_16525024","title":"JRAB/MICAL-L2 is a junctional Rab13-binding protein mediating the endocytic recycling of occludin.","date":"2006","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/16525024","citation_count":84,"is_preprint":false},{"pmid":"20008558","id":"PMC_20008558","title":"Rab13 regulates neurite outgrowth in PC12 cells through its effector protein, JRAB/MICAL-L2.","date":"2009","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/20008558","citation_count":68,"is_preprint":false},{"pmid":"26538022","id":"PMC_26538022","title":"A complex of Rab13 with MICAL-L2 and α-actinin-4 is essential for insulin-dependent GLUT4 exocytosis.","date":"2015","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/26538022","citation_count":52,"is_preprint":false},{"pmid":"18332111","id":"PMC_18332111","title":"Involvement of actinin-4 in the recruitment of JRAB/MICAL-L2 to cell-cell junctions and the formation of functional tight junctions.","date":"2008","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/18332111","citation_count":48,"is_preprint":false},{"pmid":"25864591","id":"PMC_25864591","title":"Silencing of MICAL-L2 suppresses malignancy of ovarian cancer by inducing mesenchymal-epithelial transition.","date":"2015","source":"Cancer letters","url":"https://pubmed.ncbi.nlm.nih.gov/25864591","citation_count":42,"is_preprint":false},{"pmid":"23100251","id":"PMC_23100251","title":"Rab13 small G protein and junctional Rab13-binding protein (JRAB) orchestrate actin cytoskeletal organization during epithelial junctional development.","date":"2012","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/23100251","citation_count":37,"is_preprint":false},{"pmid":"31034158","id":"PMC_31034158","title":"MICAL-L2 potentiates Cdc42-dependent EGFR stability and promotes gastric cancer cell migration.","date":"2019","source":"Journal of cellular and molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/31034158","citation_count":33,"is_preprint":false},{"pmid":"17891173","id":"PMC_17891173","title":"Involvement of Rab13 and JRAB/MICAL-L2 in epithelial cell scattering.","date":"2007","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/17891173","citation_count":31,"is_preprint":false},{"pmid":"28416812","id":"PMC_28416812","title":"A new locus regulating MICALL2 expression was identified for association with executive inhibition in children with attention deficit hyperactivity disorder.","date":"2017","source":"Molecular psychiatry","url":"https://pubmed.ncbi.nlm.nih.gov/28416812","citation_count":22,"is_preprint":false},{"pmid":"38203692","id":"PMC_38203692","title":"Comprehensive Analysis of MICALL2 Reveals Its Potential Roles in EGFR Stabilization and Ovarian Cancer Cell Invasion.","date":"2023","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/38203692","citation_count":20,"is_preprint":false},{"pmid":"27582384","id":"PMC_27582384","title":"Conformational plasticity of JRAB/MICAL-L2 provides \"law and order\" in collective cell migration.","date":"2016","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/27582384","citation_count":18,"is_preprint":false},{"pmid":"23890175","id":"PMC_23890175","title":"Junctional Rab13-binding protein (JRAB) regulates cell spreading via filamins.","date":"2013","source":"Genes to cells : devoted to molecular & cellular mechanisms","url":"https://pubmed.ncbi.nlm.nih.gov/23890175","citation_count":15,"is_preprint":false},{"pmid":"36307841","id":"PMC_36307841","title":"MICALL2 as a substrate of ubiquitinase TRIM21 regulates tumorigenesis of colorectal cancer.","date":"2022","source":"Cell communication and signaling : CCS","url":"https://pubmed.ncbi.nlm.nih.gov/36307841","citation_count":14,"is_preprint":false},{"pmid":"31488862","id":"PMC_31488862","title":"Actin Cytoskeletal Reorganization Function of JRAB/MICAL-L2 Is Fine-tuned by Intramolecular Interaction between First LIM Zinc Finger and C-terminal Coiled-coil Domains.","date":"2019","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/31488862","citation_count":13,"is_preprint":false},{"pmid":"33520979","id":"PMC_33520979","title":"MICAL-L2 Is Essential for c-Myc Deubiquitination and Stability in Non-small Cell Lung Cancer Cells.","date":"2021","source":"Frontiers in cell and developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/33520979","citation_count":11,"is_preprint":false},{"pmid":"18413246","id":"PMC_18413246","title":"Identification and characterization of JRAB/MICAL-L2, a junctional Rab13-binding protein.","date":"2008","source":"Methods in enzymology","url":"https://pubmed.ncbi.nlm.nih.gov/18413246","citation_count":8,"is_preprint":false},{"pmid":"37690696","id":"PMC_37690696","title":"MICALL2 participates in the regulation of epithelial-mesenchymal transition in alveolar epithelial cells - Potential roles in pulmonary fibrosis.","date":"2023","source":"Archives of biochemistry and biophysics","url":"https://pubmed.ncbi.nlm.nih.gov/37690696","citation_count":6,"is_preprint":false},{"pmid":"29468157","id":"PMC_29468157","title":"Dancing Styles of Collective Cell Migration: Image-Based Computational Analysis of JRAB/MICAL-L2.","date":"2018","source":"Frontiers in cell and developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/29468157","citation_count":5,"is_preprint":false},{"pmid":"38729448","id":"PMC_38729448","title":"MICAL-L2, as an estrogen-responsive gene, is involved in ER-positive breast cancer cell progression and tamoxifen sensitivity via the AKT/mTOR pathway.","date":"2024","source":"Biochemical pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/38729448","citation_count":5,"is_preprint":false},{"pmid":"36290165","id":"PMC_36290165","title":"Copy Number Variations in the MICALL2 and MOGAT2 Genes Are Associated with Ashidan Yak Growth Traits.","date":"2022","source":"Animals : an open access journal from MDPI","url":"https://pubmed.ncbi.nlm.nih.gov/36290165","citation_count":4,"is_preprint":false},{"pmid":"36811029","id":"PMC_36811029","title":"Micall2 Is Responsible for the Malignancy of Clear Cell Renal Cell Carcinoma.","date":"2023","source":"Yonago acta medica","url":"https://pubmed.ncbi.nlm.nih.gov/36811029","citation_count":3,"is_preprint":false},{"pmid":"39689763","id":"PMC_39689763","title":"High MICAL-L2 promotes cancer progression and drug resistance in renal clear cell carcinoma cells through stabilization of ACTN4 following vimentin expression.","date":"2024","source":"Biochimica et biophysica acta. Molecular basis of disease","url":"https://pubmed.ncbi.nlm.nih.gov/39689763","citation_count":2,"is_preprint":false},{"pmid":"41617093","id":"PMC_41617093","title":"MICALL2 promotes angiogenesis of colorectal cancer by regulating the EGFR/PI3K/AKT/KLF5/VEGFA axis.","date":"2026","source":"Biochemical pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/41617093","citation_count":0,"is_preprint":false},{"pmid":"41461119","id":"PMC_41461119","title":"IMB16-5 alleviates liver fibrosis by targeting MICALL2/β-catenin signaling.","date":"2025","source":"International immunopharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/41461119","citation_count":0,"is_preprint":false},{"pmid":"42174657","id":"PMC_42174657","title":"DUSP4-mediated dephosphorylation of PSMD14 enhances PSMD14-dependent deubiquitination of MICALL2 and promotes malignant progression in clear cell renal cell carcinoma.","date":"2026","source":"Journal of translational medicine","url":"https://pubmed.ncbi.nlm.nih.gov/42174657","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.09.03.674114","title":"Probing the proteome-wide impact of inhibitors of Leucine-rich Repeat Kinases 1 and 2 on protein-protein interactions and phosphorylation","date":"2025-09-07","source":"bioRxiv","url":"https://doi.org/10.1101/2025.09.03.674114","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":16195,"output_tokens":5718,"usd":0.067178,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":14251,"output_tokens":5231,"usd":0.101015,"stage2_stop_reason":"end_turn"},"total_usd":0.168193,"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\": 2006,\n      \"finding\": \"MICAL-L2 (JRAB) was identified as a Rab13 effector protein that specifically binds the GTP-bound form of Rab13 via its C-terminal coiled-coil domain, localizes at tight junctions in epithelial cells, and mediates endocytic recycling of occludin (but not transferrin receptor). A MICAL-L2 mutant lacking the Rab13-binding domain (MICAL-L2-N) specifically inhibited occludin recycling and blocked TJ formation in Ca2+-switch assays. MICAL-L2 was displaced from TJs upon actin depolymerization and redistributed along actin structures, indicating it links Rab13 to the actin cytoskeleton.\",\n      \"method\": \"Immunoprecipitation, immunofluorescence, recycling assay, Ca2+-switch assay, yeast two-hybrid (initial identification), dominant-negative mutant expression\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, multiple orthogonal functional assays (recycling, Ca2+-switch, dominant-negative), replicated in subsequent studies\",\n      \"pmids\": [\"16525024\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"JRAB/MICAL-L2 interacts with both Rab8 and Rab13 via its C-terminal domain, and these two Rab GTPases compete with each other for MICAL-L2 binding. Rab8 and Rab13 functionally associate with MICAL-L2 at perinuclear recycling/storage compartments and at the plasma membrane, respectively. Rab13 knockdown suppressed claudin-1 and occludin but not E-cadherin transport, while Rab8 knockdown inhibited Rab13-independent E-cadherin transport; MICAL-L2 knockdown or dominant-negative MICAL-L2-C expression inhibited transport of all three junctional proteins, coordinating TJ and AJ assembly.\",\n      \"method\": \"siRNA knockdown, dominant-negative mutant expression, Ca2+-switch assay, immunofluorescence, co-immunoprecipitation\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (KD, dominant-negative, Co-IP, imaging), builds on prior replicated findings\",\n      \"pmids\": [\"18094055\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"Actinin-4 was identified as a binding partner for the plasma membrane-targeting domain of JRAB/MICAL-L2 using yeast two-hybrid and confirmed by co-immunoprecipitation. Actinin-4 colocalizes with JRAB/MICAL-L2 at cell-cell junctions and links JRAB/MICAL-L2 to F-actin. The actinin-4–MICAL-L2 interaction was enhanced by Rab13 activation. siRNA depletion of actinin-4 delayed JRAB/MICAL-L2 recruitment to cell-cell junctions and impaired functional TJ formation during epithelial polarization.\",\n      \"method\": \"Yeast two-hybrid, co-immunoprecipitation, siRNA knockdown, Ca2+-switch assay, immunofluorescence\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — yeast two-hybrid plus reciprocal Co-IP plus siRNA functional validation, multiple orthogonal methods\",\n      \"pmids\": [\"18332111\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"JRAB/MICAL-L2 domain structure was characterized: it contains a calponin-homology (CH) domain at the N-terminus, a LIM domain in the middle, and a coiled-coil domain at the C-terminus. The C-terminus specifically binds the GTP-bound form of Rab13. In epithelial cells it localizes to tight junctions; in fibroblasts it distributes along stress fibers.\",\n      \"method\": \"Domain deletion analysis, immunoprecipitation, immunofluorescence, yeast two-hybrid (referenced prior work)\",\n      \"journal\": \"Methods in enzymology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — consolidates prior structural characterization data, single review/methods paper but based on earlier experimental work\",\n      \"pmids\": [\"18413246\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"JRAB/MICAL-L2 exhibits an intramolecular interaction between its N-terminal CH and LIM domains and the C-terminal coiled-coil domain. Rab13 binding to JRAB/MICAL-L2 stimulates interaction of MICAL-L2 with actinin-4, which localizes to cell body and neurite tips in PC12 cells. Expression of MICAL-L2 alone inhibits neurite outgrowth, but co-expression of dominant-active Rab13 rescues this phenotype, suggesting Rab13-dependent conformational change enables transfer of actinin-4 to neurite tips to reorganize the actin cytoskeleton.\",\n      \"method\": \"Overexpression/dominant-active mutants, co-immunoprecipitation, immunofluorescence, neurite outgrowth assay in PC12 cells\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — intramolecular interaction mapped by deletion/co-IP, functional rescue with dominant-active Rab13, multiple orthogonal methods in single study\",\n      \"pmids\": [\"20008558\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"Rab13 and JRAB/MICAL-L2 are involved in epithelial cell scattering in response to TPA. During TPA-induced MDCK cell scattering, Rab13 was transiently activated, and both Rab13 and JRAB/MICAL-L2 co-localized with F-actin at cell-cell contact sites before accumulating at lamellipodia. Knockdown of JRAB/MICAL-L2 inhibited TPA-induced scattering, rescued by re-expression of mouse JRAB/MICAL-L2.\",\n      \"method\": \"siRNA knockdown, rescue by re-expression, immunofluorescence, Rab13 activation assay\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — siRNA KD with rescue experiment, colocalization, single lab\",\n      \"pmids\": [\"17891173\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"JRAB/MICAL-L2 interacts with both actinin-1 and actinin-4 and with filamentous actin (F-actin) via different domains, and regulates actin cross-linking and stabilization. During epithelial junctional development, JRAB is enriched in the actin bundle at the free border and undergoes a Rab13-dependent conformational change required for maturation of cell-cell adhesion.\",\n      \"method\": \"Co-immunoprecipitation, domain deletion analysis, immunofluorescence, live imaging, Rab13 dominant-active/dominant-negative mutants\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple binding partners mapped with Co-IP/domain analysis, Rab13-dependent conformational change demonstrated, consistent with prior replicated findings\",\n      \"pmids\": [\"23100251\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"In the 'open' conformation (JRABΔCC mutant), JRAB/MICAL-L2 co-immunoprecipitates with filamin (an actin cross-linking protein). Expression of ASB2, which induces degradation of all three filamin isoforms, inhibited JRABΔCC-induced cell spreading with membrane ruffles, demonstrating that JRAB regulates actin cytoskeletal reorganization and cell spreading through filamins. In contrast, actinin-1/-4 co-precipitated with JRABΔCC but did not affect cell spreading.\",\n      \"method\": \"Co-immunoprecipitation with conformational mutants, ASB2-mediated filamin degradation, morphological analysis in NIH3T3 cells\",\n      \"journal\": \"Genes to cells\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP with functional validation via filamin depletion, single lab, two methods\",\n      \"pmids\": [\"23890175\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Rab13 engages MICAL-L2 as a scaffold upon insulin stimulation in muscle cells. Insulin increased Rab13 binding to MICAL-L2 and binding of MICAL-L2 to α-actinin-4 (ACTN4), forming an insulin-dependent ternary complex (Rab13–MICAL-L2–ACTN4). GLUT4 associated with this complex in response to insulin via the ACTN4-binding domain of MICAL-L2. Knockdown of MICAL-L2 or expression of truncated MICAL-L2-CT impaired insulin-stimulated GLUT4 translocation to the plasma membrane.\",\n      \"method\": \"Co-immunoprecipitation, pull-down with MICAL-L2 fragments, siRNA knockdown, confocal fluorescence, TIRF microscopy, structured illumination microscopy\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (pull-down, Co-IP, TIRF/SIM imaging, siRNA KD), domain mapping of ACTN4 interaction, consistent functional readout\",\n      \"pmids\": [\"26538022\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"JRAB/MICAL-L2 undergoes a Rab13-dependent conformational change between closed and open states. A structural model was generated by bioinformatic and biochemical analyses. Impairment of MICAL-L2 conformational plasticity (using fixed conformation mutants) caused excessive rigidity and loss of directionality in collective cell migration. Live imaging and computational analysis showed that closed/open conformations dictate distinct collective migration behaviors.\",\n      \"method\": \"Structural modeling, biochemical conformation assays, live imaging, computational biomechanics, mutant expression\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — structural model validated biochemically, functional consequence of conformational change demonstrated by live imaging and quantitative cell biology; multiple orthogonal methods\",\n      \"pmids\": [\"27582384\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"The first LIM zinc finger domain of JRAB/MICAL-L2 binds the first and second actin molecules at the minus end of F-actin, potentially inhibiting actin filament depolymerization. This same zinc finger domain also contributes to the intramolecular interaction with the C-terminal coiled-coil domain, and residues responsible for intramolecular interaction overlap with those involved in F-actin association, providing a mechanism for fine-tuning JRAB function.\",\n      \"method\": \"Biochemical actin-binding assays, mutagenesis, intramolecular interaction mapping, co-immunoprecipitation with mutants\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — actin-binding assay with mutagenesis, intramolecular interaction mapped, single lab\",\n      \"pmids\": [\"31488862\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"MICAL-L2 expression upregulated EGFR protein level through a transcription-independent mechanism involving inhibition of EGFR degradation in lysosomes. This effect on EGFR stability was mediated in a Cdc42-dependent manner, activating downstream HSP27/cytoskeleton and HSP27/β-catenin signaling pathways to promote gastric cancer cell migration.\",\n      \"method\": \"siRNA knockdown, overexpression, Western blotting, lysosome inhibitor experiments, Cdc42 activation assays, immunofluorescence\",\n      \"journal\": \"Journal of cellular and molecular medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — single lab, multiple assays but mechanistic basis of Cdc42 dependence not fully reconstituted in vitro\",\n      \"pmids\": [\"31034158\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"MICAL-L2 physically interacts with c-Myc protein (co-immunoprecipitation), maintains nuclear c-Myc levels, prolongs its half-life, and inhibits c-Myc polyubiquitination. MICAL-L2 knockdown accelerated c-Myc degradation through polyubiquitylation; MICAL-L2 blocked c-Myc phosphorylation at Thr58, which is required for ubiquitin-dependent c-Myc degradation, thereby acting as a deubiquitination/stabilization factor for c-Myc in NSCLC cells.\",\n      \"method\": \"Co-immunoprecipitation, immunofluorescence, polyubiquitylation detection assay, protein stability assay, siRNA knockdown, Western blotting\",\n      \"journal\": \"Frontiers in cell and developmental biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP confirmed interaction, ubiquitination assays and Thr58 phosphorylation mechanistically linked, single lab\",\n      \"pmids\": [\"33520979\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"TRIM21 (ubiquitin E3 ligase) mediates ubiquitination and proteasome-dependent degradation of MICALL2, acting as a negative regulator. Co-immunoprecipitation and mass spectrometry identified TRIM21 as a MICALL2-interacting protein. TRIM21 expression negatively correlates with MICALL2 levels and reversely regulates MICALL2-dependent tumorigenic activity (growth and migration via Wnt/β-catenin) in colorectal cancer cells.\",\n      \"method\": \"Co-immunoprecipitation, mass spectrometry, ubiquitination assays, siRNA/overexpression functional assays, Western blotting\",\n      \"journal\": \"Cell communication and signaling\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP plus mass spectrometry identification, ubiquitination assay confirms TRIM21 as E3 ligase, single lab\",\n      \"pmids\": [\"36307841\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"MICALL2 silencing in ovarian cancer cells promoted EGFR lysosomal degradation in a Rac1-dependent (rather than Cdc42-dependent as in gastric cancer) manner. Rac1 suppression attenuated MICALL2-induced pro-EGFR, pro-MMP9, and pro-invasion effects. MMP9 was identified as the target gene downstream of MICALL2 via EGFR-AKT-mTOR signaling, regulating invadopodium formation and cell invasion.\",\n      \"method\": \"siRNA knockdown, overexpression, Rac1 inhibition, autophagy inhibitors (acadesine, chloroquine), co-immunoprecipitation, invasion assays\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — mechanistic pathway defined by pharmacological inhibition and KD, single lab, no in vitro reconstitution\",\n      \"pmids\": [\"38203692\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"MICAL-L2 stabilizes ACTN4 protein in a Rab13-dependent manner in clear cell renal cell carcinoma (KIRC) cells, reducing ACTN4 degradation. This MICAL-L2–ACTN4 complex interaction was confirmed by co-immunoprecipitation. ACTN4 stabilization leads to increased Vimentin expression, promoting cancer progression and resistance to sunitinib and everolimus.\",\n      \"method\": \"Co-immunoprecipitation, siRNA knockdown, overexpression, Western blotting, wound-healing, Transwell, CCK-8 assays\",\n      \"journal\": \"Biochimica et biophysica acta. Molecular basis of disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — Co-IP confirmed MICAL-L2/ACTN4 interaction with Rab13 dependence, functional assays, single lab\",\n      \"pmids\": [\"39689763\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"MICALL2 prevents β-catenin from binding to GSK3β, thereby shielding β-catenin from degradation and promoting Wnt/β-catenin signaling in hepatic stellate cells. A small molecule IMB16-5 binds to the C-terminal domain of MICALL2 (identified by LiP-SMap and molecular dynamics simulation) and disrupts the MICALL2–β-catenin interaction, suppressing β-catenin nuclear translocation and HSC activation. MICALL2 overexpression in vivo abolished the therapeutic efficacy of IMB16-5, confirming target engagement.\",\n      \"method\": \"LiP-SMap (limited proteolysis-small molecule mapping), molecular dynamics simulation, co-immunoprecipitation, in vivo mouse fibrosis model, overexpression rescue experiment\",\n      \"journal\": \"International immunopharmacology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — LiP-SMap maps drug binding site, Co-IP shows MICALL2-β-catenin-GSK3β relationship, in vivo confirmation, single lab\",\n      \"pmids\": [\"41461119\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"PSMD14 (a proteasome deubiquitinase) physically interacts with MICALL2 (confirmed by co-immunoprecipitation) and suppresses MICALL2 ubiquitination and degradation, thereby increasing MICALL2 protein levels. DUSP4-mediated dephosphorylation of PSMD14 enhances PSMD14–MICALL2 interaction and amplifies PSMD14's stabilizing effect on MICALL2, promoting ccRCC malignant progression.\",\n      \"method\": \"Co-immunoprecipitation, immunoprecipitation for ubiquitination analysis, PSMD14 phosphorylation analysis, siRNA knockdown, Western blotting, in vivo xenograft model\",\n      \"journal\": \"Journal of translational medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP confirmed PSMD14–MICALL2 interaction, ubiquitination and phosphorylation assays, functional validation in vivo, single lab\",\n      \"pmids\": [\"42174657\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"MICALL2 was identified as a candidate substrate of LRRK2 kinase in proximity-labeling mass spectrometry and quantitative phosphoproteomics experiments. LRRK2 inhibitors altered MICALL2 phosphorylation status at the proteome-wide level.\",\n      \"method\": \"Proximity labeling mass spectrometry (BioID/TurboID), quantitative phosphoproteomics, LRRK1/2 inhibitor treatment\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 4 / Weak — identified as candidate substrate in large-scale screen, no direct in vitro kinase assay or site-specific mutagenesis confirming LRRK2-mediated phosphorylation of MICALL2\",\n      \"pmids\": [\"bio_10.1101_2025.09.03.674114\"],\n      \"is_preprint\": true\n    }\n  ],\n  \"current_model\": \"MICALL2/JRAB is a multidomain scaffold protein (CH–LIM–coiled-coil) that functions as an effector of GTP-bound Rab13 (and Rab8) to coordinate vesicle trafficking and actin cytoskeletal reorganization: in the Rab13-bound open conformation it recruits actinin-1/4 and filamins to regulate actin cross-linking and stabilization, mediates endocytic recycling of tight junction proteins (occludin, claudins) and E-cadherin to the plasma membrane, undergoes a Rab13-dependent intramolecular conformational switch that controls collective cell migration directionality, and also acts in non-junctional contexts to stabilize EGFR (via Cdc42/Rac1-dependent inhibition of lysosomal degradation), stabilize c-Myc (by blocking Thr58 phosphorylation and polyubiquitination), protect β-catenin from GSK3β-mediated degradation to activate Wnt signaling, and stabilize ACTN4 in a Rab13-dependent manner; its own protein stability is regulated by TRIM21-mediated ubiquitination/proteasomal degradation and by PSMD14-mediated deubiquitination.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"MICALL2 (JRAB) is a multidomain scaffold protein (CH–LIM–coiled-coil) that acts as an effector of GTP-bound Rab13, and also Rab8, to couple vesicle trafficking with actin cytoskeletal reorganization at epithelial junctions [#0, #1, #3]. Through its C-terminal coiled-coil it binds active Rab13/Rab8, and through its membrane-targeting and LIM regions it engages the actin cross-linkers actinin-1/4 and filamins and binds F-actin directly, thereby regulating actin cross-linking and stabilization [#2, #6, #7, #10]. In this capacity it mediates endocytic recycling of tight- and adherens-junction proteins—occludin, claudin-1, and E-cadherin—to the plasma membrane and is required for coordinated TJ/AJ assembly [#0, #1]. MICALL2 operates as a conformational switch: an intramolecular interaction between its N-terminal CH/LIM domains and the C-terminal coiled-coil holds it closed, and Rab13 binding opens the molecule to transfer actinin-4 onto actin structures; this Rab13-dependent open/closed plasticity governs actin remodeling, junctional maturation, and the directionality of collective cell migration [#4, #6, #9, #10]. Beyond junctions, MICALL2 serves as a Rab13-dependent scaffold linking GLUT4 to a Rab13–MICALL2–ACTN4 ternary complex during insulin-stimulated GLUT4 translocation [#8]. In cancer contexts MICALL2 acts as a stability factor for multiple oncogenic effectors—stabilizing EGFR by blocking its lysosomal degradation via Cdc42- or Rac1-dependent signaling [#11, #14], stabilizing c-Myc by preventing Thr58 phosphorylation and polyubiquitination [#12], shielding β-catenin from GSK3β to activate Wnt signaling [#16], and stabilizing ACTN4 in a Rab13-dependent manner [#15]. MICALL2's own abundance is set by opposing ubiquitin machinery: TRIM21 ubiquitinates it for proteasomal degradation, while the proteasome-associated deubiquitinase PSMD14 stabilizes it [#13, #17].\",\n  \"teleology\": [\n    {\n      \"year\": 2006,\n      \"claim\": \"Established MICALL2 as a Rab13 effector that links a small GTPase to junctional vesicle trafficking, answering how Rab13 controls tight junction assembly.\",\n      \"evidence\": \"Co-IP, recycling and Ca2+-switch assays with a Rab13-binding-deficient mutant in epithelial cells\",\n      \"pmids\": [\"16525024\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not resolve the structural basis of GTP-dependent Rab13 binding\", \"Cargo selectivity (occludin vs transferrin receptor) mechanism unexplained\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Showed MICALL2 integrates two Rab GTPases by competitive binding, explaining how it coordinates both tight- and adherens-junction protein delivery.\",\n      \"evidence\": \"siRNA knockdown, dominant-negative MICAL-L2-C, Ca2+-switch and Co-IP in epithelial cells\",\n      \"pmids\": [\"18094055\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not define how Rab8 vs Rab13 compartment selection is regulated\", \"Spatial control of competition not addressed\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Linked MICALL2/Rab13 to epithelial scattering, extending its role from static junctions to migratory behavior.\",\n      \"evidence\": \"siRNA knockdown with re-expression rescue and Rab13 activation assay in TPA-stimulated MDCK cells\",\n      \"pmids\": [\"17891173\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Did not identify the molecular trigger coupling Rab13 activation to scattering\", \"Single-lab observation\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Identified actinin-4 as the link between MICALL2 and F-actin and defined the CH–LIM–coiled-coil domain architecture, providing the structural framework for its scaffold function.\",\n      \"evidence\": \"Yeast two-hybrid, Co-IP, siRNA, domain-deletion analysis and immunofluorescence\",\n      \"pmids\": [\"18332111\", \"18413246\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Precise actinin-4-binding domain boundaries partially defined\", \"How Rab13 enhances the actinin-4 interaction left mechanistically open\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Discovered the Rab13-driven intramolecular conformational switch, explaining how Rab13 binding mobilizes actinin-4 to remodel actin.\",\n      \"evidence\": \"Co-IP, dominant-active Rab13, neurite-outgrowth rescue in PC12 cells\",\n      \"pmids\": [\"20008558\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Atomic structure of open vs closed states not determined\", \"Kinetics of the conformational transition unknown\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Demonstrated MICALL2 binds actinin-1/4 and F-actin through distinct domains to control actin cross-linking and junctional maturation, consolidating its cytoskeletal-regulatory role.\",\n      \"evidence\": \"Co-IP, domain deletion, live imaging with Rab13 mutants\",\n      \"pmids\": [\"23100251\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Quantitative effect on actin cross-linking not measured in vitro\", \"Hierarchy of multiple actin-binding partners unresolved\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Showed the open conformation recruits filamins to drive cell spreading, distinguishing functional outputs of different MICALL2-bound cross-linkers.\",\n      \"evidence\": \"Co-IP with conformational mutants and ASB2-mediated filamin degradation in NIH3T3 cells\",\n      \"pmids\": [\"23890175\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Why actinins co-precipitate but do not affect spreading unexplained\", \"Single-lab, two-method support\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Extended the scaffold function to metabolic trafficking by placing MICALL2 in an insulin-induced Rab13–MICALL2–ACTN4 complex that delivers GLUT4.\",\n      \"evidence\": \"Pull-down, Co-IP, siRNA, TIRF/SIM imaging in muscle cells\",\n      \"pmids\": [\"26538022\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How insulin signaling triggers Rab13–MICALL2 binding not defined\", \"Direct GLUT4 contact vs indirect not resolved\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Built a structural model of the conformational switch and tied conformational plasticity directly to collective migration directionality.\",\n      \"evidence\": \"Structural modeling, biochemical conformation assays, live imaging and computational biomechanics with fixed-conformation mutants\",\n      \"pmids\": [\"27582384\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Model not validated by experimental high-resolution structure\", \"Upstream control of conformational state during migration unclear\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Defined a direct F-actin-binding mechanism for the LIM zinc finger and showed overlap with the intramolecular interface, providing a molecular basis for fine-tuning MICALL2 activity.\",\n      \"evidence\": \"Biochemical actin-binding assays, mutagenesis, intramolecular interaction mapping\",\n      \"pmids\": [\"31488862\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional consequence of minus-end binding on filament dynamics not tested in cells\", \"Single-lab biochemistry\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Opened a non-junctional oncogenic role by showing MICALL2 stabilizes EGFR via Cdc42-dependent inhibition of lysosomal degradation.\",\n      \"evidence\": \"Knockdown/overexpression, lysosome inhibitors, Cdc42 activation assays in gastric cancer cells\",\n      \"pmids\": [\"31034158\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct biochemical link between MICALL2 and EGFR trafficking not reconstituted\", \"Cdc42-dependence mechanism incomplete\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Identified MICALL2 as a c-Myc stabilizer that blocks Thr58 phosphorylation and polyubiquitination, broadening its function to nuclear oncoprotein regulation.\",\n      \"evidence\": \"Co-IP, polyubiquitylation and protein-stability assays, siRNA in NSCLC cells\",\n      \"pmids\": [\"33520979\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether the interaction is direct or how it blocks Thr58 phosphorylation unresolved\", \"Single-lab\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Defined upstream control of MICALL2 abundance by identifying TRIM21 as its degradative E3 ligase.\",\n      \"evidence\": \"Co-IP, mass spectrometry, ubiquitination assays and functional readouts in colorectal cancer cells\",\n      \"pmids\": [\"36307841\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"TRIM21 ubiquitination site(s) on MICALL2 not mapped\", \"Single-lab\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Showed EGFR stabilization can proceed through Rac1 rather than Cdc42, with MMP9 as a downstream effector, indicating context-dependent GTPase usage.\",\n      \"evidence\": \"Knockdown/overexpression, Rac1 inhibition, autophagy inhibitors, invasion assays in ovarian cancer cells\",\n      \"pmids\": [\"38203692\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Basis for tissue-specific Cdc42 vs Rac1 choice unexplained\", \"No in vitro reconstitution\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Demonstrated Rab13-dependent stabilization of ACTN4 links MICALL2 to vimentin upregulation and therapy resistance, connecting its scaffold role to malignant progression.\",\n      \"evidence\": \"Co-IP, siRNA/overexpression and functional assays in clear cell renal cell carcinoma cells\",\n      \"pmids\": [\"39689763\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism by which MICALL2 protects ACTN4 from degradation undefined\", \"Single-lab\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Established that MICALL2 shields β-catenin from GSK3β to activate Wnt signaling and identified a small molecule binding its C-terminus that disrupts this function.\",\n      \"evidence\": \"LiP-SMap, molecular dynamics, Co-IP and in vivo fibrosis model with overexpression rescue\",\n      \"pmids\": [\"41461119\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct vs scaffolded MICALL2–β-catenin contact not fully resolved\", \"Single-lab\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Completed the regulatory loop on MICALL2 stability by showing the deubiquitinase PSMD14, controlled by DUSP4-mediated dephosphorylation, stabilizes MICALL2.\",\n      \"evidence\": \"Co-IP, ubiquitination and phosphorylation assays, siRNA, in vivo xenograft in ccRCC\",\n      \"pmids\": [\"42174657\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether TRIM21 and PSMD14 act on the same ubiquitin sites unknown\", \"Single-lab\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Flagged MICALL2 as a candidate LRRK2 kinase substrate, raising the possibility of phosphoregulation of the scaffold.\",\n      \"evidence\": \"Proximity-labeling MS and quantitative phosphoproteomics with LRRK inhibitors (preprint)\",\n      \"pmids\": [\"bio_10.1101_2025.09.03.674114\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No direct in vitro kinase assay or site-specific mutagenesis confirming LRRK2-mediated phosphorylation\", \"Functional consequence of phosphorylation unknown\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How the Rab13-driven conformational switch is mechanistically coupled to MICALL2's diverse protein-stabilization activities (EGFR, c-Myc, β-catenin, ACTN4) remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No high-resolution structure of full-length MICALL2 in either conformation\", \"Whether stabilization activities are direct or scaffold-dependent unestablished\", \"Upstream signals selecting junctional vs oncogenic functions undefined\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 2, 6, 8]},\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [2, 6, 7, 10]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [11, 12, 15, 16]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [0, 1, 8]},\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [0, 3, 6]},\n      {\"term_id\": \"GO:0031410\", \"supporting_discovery_ids\": [0, 1]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-5653656\", \"supporting_discovery_ids\": [0, 1, 8]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [11, 16]},\n      {\"term_id\": \"R-HSA-1500931\", \"supporting_discovery_ids\": [0, 1, 2]}\n    ],\n    \"complexes\": [\"Rab13–MICALL2–ACTN4 ternary complex\"],\n    \"partners\": [\"RAB13\", \"RAB8A\", \"ACTN4\", \"ACTN1\", \"TRIM21\", \"PSMD14\", \"MYC\", \"CTNNB1\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":7,"faith_total":7,"faith_pct":100.0}}