{"gene":"MAPRE2","run_date":"2026-06-10T02:59:50","timeline":{"discoveries":[{"year":2016,"finding":"Aurora B and CDK1 phosphorylate MAPRE2/EB2 at multiple sites within the amino terminus and a cluster of serine/threonine residues in the linker connecting the calponin homology and end-binding homology domains during mitotic entry and progression. This phosphorylation reduces MAPRE2 binding affinity for microtubules. Expression of non-phosphorylatable MAPRE2 stabilizes kinetochore microtubule dynamics, delays bipolar metaphase plate formation, and leads to aneuploidy.","method":"In vitro kinase assays, phospho-site mutagenesis (non-phosphorylatable mutants), live-cell imaging, kinetochore microtubule dynamics assays","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro kinase assay with mutagenesis plus defined cellular phenotype in single rigorous study","pmids":["27030108"],"is_preprint":false},{"year":2013,"finding":"MAPRE2/EB2 is required for initial microtubule reorganisation during apico-basal epithelial differentiation. siRNA knockdown of EB2 in undifferentiated epithelial cells induced formation of straight, less dynamic microtubules with EB1 and ACF7 lattice association and co-alignment with actin filaments, a phenotype rescued by formin inhibition. Conversely, EB2 downregulation at later differentiation stages promoted microtubule stability and bundle formation.","method":"siRNA knockdown, live-cell imaging, immunofluorescence, formin inhibitor rescue experiments, in situ tissue analysis","journal":"Journal of cell science","confidence":"High","confidence_rationale":"Tier 2 / Moderate — siRNA knockdown with defined cellular phenotype, rescue experiment, and orthogonal in situ tissue validation","pmids":["23813963"],"is_preprint":false},{"year":2015,"finding":"MAPRE2 mutations associated with circumferential skin creases Kunze type (CSC-KT) syndrome result in enhanced MAPRE2 binding to microtubules, implying an increased dwell time at microtubule plus ends. In a zebrafish craniofacial development model, MAPRE2 mutations perturb branchial arch patterning.","method":"Microtubule co-sedimentation/binding assays with patient-derived mutants, zebrafish in vivo craniofacial analysis","journal":"American journal of human genetics","confidence":"High","confidence_rationale":"Tier 1-2 / Moderate — direct biochemical binding assay with mutants plus in vivo zebrafish model, single study with two orthogonal methods","pmids":["26637975"],"is_preprint":false},{"year":2015,"finding":"MAPRE2/EB2 interacts specifically with HAX1 (HCLS1-associated protein X-1), identified by quantitative proteomics. Knockdown of HAX1 or EB2 in skin epidermal cells stabilizes focal adhesions and impairs epidermal migration in vitro and in vivo. The EB2-HAX1 interaction is required for focal adhesion turnover and cell motility.","method":"Quantitative proteomics (mass spectrometry), Co-immunoprecipitation, siRNA knockdown, in vitro migration assays, in vivo wound healing model","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Moderate — MS-identified interaction validated by Co-IP, loss-of-function with defined phenotype in vitro and in vivo","pmids":["26527684"],"is_preprint":false},{"year":2021,"finding":"MAPRE2/EB2 mediates microtubule destabilization, increases Src kinase activity, and thereby facilitates ERK signaling activation in hepatocellular carcinoma cells. ERK signaling in turn promotes EB2 expression, forming a positive feedback loop. ERK inhibitor U0126 blocked EB2-mediated proliferation and metastasis.","method":"Overexpression and siRNA knockdown, in vitro invasion assays, in vivo xenograft, kinase activity assays, ERK inhibitor treatment","journal":"Clinical science","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — defined pathway placement with in vitro and in vivo data but single lab, mechanism of Src activation by EB2 not directly demonstrated","pmids":["33755094"],"is_preprint":false},{"year":2020,"finding":"WDHD1 (a ubiquitin ligase) interacts with MAPRE2 in the nucleus and promotes MAPRE2 ubiquitination, leading to MAPRE2 degradation and contributing to cisplatin resistance in lung adenocarcinoma cells.","method":"Co-immunoprecipitation, protein stability assays, ubiquitination assays, immunofluorescence, MAPRE2 and WDHD1 knockout/knockdown in cell lines and animal experiments","journal":"Frontiers in oncology","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — Co-IP and ubiquitination assays with functional validation, single lab","pmids":["32426268"],"is_preprint":false},{"year":2024,"finding":"E3 ubiquitin ligase ASB14 promotes MAPRE2 ubiquitination and degradation. Loss of ASB14 decreases MAPRE2 protein degradation, promotes cardiomyocyte nuclear proliferation, and enhances cardiac repair after myocardial infarction.","method":"Microarray analysis, ASB14 silencing in vitro and in vivo, Western blot for MAPRE2 protein levels, cardiomyocyte proliferation assays","journal":"Cell biochemistry and biophysics","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — ubiquitination pathway placement with in vitro and in vivo data, single lab, mechanistic detail limited in abstract","pmids":["38319584"],"is_preprint":false},{"year":2021,"finding":"MAPRE2 mutations cause altered cranial neural crest cell migration in vitro and in vivo (xenotransplantation into chicken embryos). Changes in focal adhesion dynamics underlie the altered cell motility of MAPRE2 mutant cranial neural crest cells.","method":"Patient-derived iPSC lines with CRISPR/Cas9 isogenic controls, neural crest differentiation, in vitro migration assays, xenotransplantation into chicken embryos, focal adhesion analysis","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — iPSC-derived disease models with isogenic controls, in vitro and in vivo validation, orthogonal methods","pmids":["33654163"],"is_preprint":false},{"year":2023,"finding":"MAPRE2 loss-of-function (CRISPR knockout in zebrafish and siRNA/KO in human iPSC-CMs) decreases voltage-gated sodium channel function (reduced sodium current density) and disrupts adherens junctions with mislocalization of mature N-cadherin, associated with decreased detyrosinated tubulin. MAPRE2 knockdown in iPSC-CMs increases microtubule growth velocity and distance. Knockdown of ttl (tubulin tyrosine ligase) in mapre2 knockout larvae rescued detyrosinated tubulin levels and ventricular action potential upstroke velocity, establishing epistasis between MAPRE2 and tubulin detyrosination in regulating sodium channel function.","method":"CRISPR/Cas9 zebrafish knockout, siRNA knockdown and KO in iPSC-CMs, patch clamping, ECG, voltage mapping, immunocytochemistry, microtubule plus-end tracking, genetic epistasis (ttl knockdown rescue)","journal":"Circulation research","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (electrophysiology, imaging, genetic epistasis rescue), in vivo and in vitro, with mechanistic pathway placement","pmids":["38095085"],"is_preprint":false},{"year":2022,"finding":"MAPRE2 is distributed along the meiotic spindle during metaphase I and II in mouse oocytes. siRNA-mediated knockdown of Mapre2 severely impairs microtubule stability, kinetochore-microtubule attachment, and chromosome alignment, activates the spindle assembly checkpoint, prevents cyclin B1 degradation, and leads to failure of chromosome segregation and first polar body extrusion.","method":"siRNA knockdown, immunofluorescence, spindle assembly checkpoint markers, cyclin B1 western blot, polar body extrusion assay","journal":"Experimental cell research","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — siRNA knockdown with multiple defined cellular readouts, single lab","pmids":["35398309"],"is_preprint":false},{"year":2019,"finding":"MAPRE2/EB2 interacts with TRPM4 via a specific EB-binding motif on TRPM4. Mutations of this motif abolish the TRPM4-EB interaction and reduce plasma membrane expression of the channel. EB1 and EB2 proteins are required for TRPM4 anterograde trafficking; their loss leads to endoplasmic reticulum-associated TRPM4 distribution and affects focal adhesion disassembly and cell invasion.","method":"Co-immunoprecipitation, mutagenesis of EB-binding motif, immunofluorescence, cell invasion assays, focal adhesion assays","journal":"FASEB journal","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — Co-IP with mutagenesis and functional consequences, but EB1 and EB2 contributions not fully separated, single lab","pmids":["31112396"],"is_preprint":false},{"year":2009,"finding":"MAPRE2 expression is significantly increased in highly nerve-invasive pancreatic cancer cells compared to less invasive cells, and changes in MAPRE2 expression result in altered actin distribution in these cells.","method":"siRNA knockdown/overexpression, immunofluorescence for actin distribution, comparative expression analysis","journal":"International journal of oncology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single method (actin redistribution by immunofluorescence) with no defined molecular mechanism, single lab","pmids":["19787265"],"is_preprint":false},{"year":2022,"finding":"Transcription factor JUND binds to the MAPRE2 promoter (shown by dual-luciferase reporter and ChIP assay) and increases MAPRE2 transcription, thereby facilitating cell proliferative and angiogenic abilities in esophageal squamous cell carcinoma.","method":"Dual-luciferase reporter assay, chromatin immunoprecipitation (ChIP), siRNA knockdown, in vitro angiogenesis and proliferation assays","journal":"Tissue & cell","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — direct promoter binding demonstrated by ChIP and reporter assay, functional rescue, single lab","pmids":["36608637"],"is_preprint":false},{"year":2026,"finding":"EB2/MAPRE2 associates with the mitotic spindle in neural progenitors. A disease-causing truncation mutation (Q152X) stalls mitosis in prometaphase, causes defects in chromosome congression, misaligned chromosomes, and p53-induced apoptosis in both mouse transgenic models and human stem cell-derived neural progenitor cultures.","method":"Transgenic mouse models, stem cell-derived neural progenitor cultures, immunofluorescence, time-lapse imaging, p53 pathway analysis","journal":"Development (Cambridge, England)","confidence":"High","confidence_rationale":"Tier 2 / Moderate — disease mutation in two orthogonal systems (mouse transgenics and human iPSC-derived cultures) with defined mitotic phenotype","pmids":["41725360"],"is_preprint":false},{"year":2024,"finding":"CDKL5 phosphorylates MAPRE2/EB2, and this phosphorylation is impaired when CDKL5 undergoes truncating mutations (S726X, R781X) that disrupt its liquid-liquid phase separation. Reduced CDKL5 LLPS correlates with decreased EB2 phosphorylation.","method":"Optogenetic LLPS induction, super-resolution imaging, electron microscopy, phosphorylation assays with truncation mutants","journal":"bioRxiv (preprint)","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — phosphorylation shown in cellular assay with disease mutants, preprint not yet peer reviewed, single lab","pmids":["bio_10.1101_2024.11.18.624084"],"is_preprint":true},{"year":2024,"finding":"Patient-derived CDD neurons (with loss-of-function CDKL5 variants) show decreased phosphorylated EB2, confirming EB2 as a molecular substrate of CDKL5 in human neurons.","method":"iPSC differentiation (NGN2-induced neurons and cortical organoids), Western blot for phospho-EB2","journal":"bioRxiv (preprint)","confidence":"Low","confidence_rationale":"Tier 3 / Weak — indirect evidence (reduced substrate phosphorylation in disease context), preprint, single readout","pmids":["bio_10.1101_2024.11.11.622878"],"is_preprint":true}],"current_model":"MAPRE2/EB2 is a microtubule plus-end tracking protein whose binding to microtubules is regulated by Aurora B- and CDK1-mediated phosphorylation during mitosis (and by CDKL5 phosphorylation in neurons), which controls kinetochore microtubule dynamics, mitotic progression, and genome stability; it promotes apico-basal epithelial differentiation by modulating microtubule reorganization and EB1/ACF7 deployment; it drives focal adhesion turnover and cell migration via an EB2–HAX1 complex; it facilitates TRPM4 anterograde trafficking to the plasma membrane; it regulates ventricular sodium channel function through a microtubule detyrosination pathway; its ubiquitination and degradation is mediated by the E3 ligases WDHD1 and ASB14; and gain-of-function mutations increase its dwell time at microtubule plus ends, perturbing neural crest migration and craniofacial development underlying CSC-KT syndrome."},"narrative":{"mechanistic_narrative":"MAPRE2 (EB2) is a microtubule plus-end tracking protein that controls microtubule dynamics across mitosis, epithelial differentiation, cell migration, and membrane protein trafficking [PMID:27030108, PMID:23813963, PMID:26527684]. During mitotic entry and progression, Aurora B and CDK1 phosphorylate MAPRE2 at the amino terminus and the linker between its calponin-homology and end-binding domains, reducing its microtubule binding affinity; blocking this phosphorylation stabilizes kinetochore microtubule dynamics, delays metaphase plate formation, and produces aneuploidy [PMID:27030108]. Consistent with a conserved role in spindle function, MAPRE2 decorates the meiotic spindle and is required for kinetochore-microtubule attachment, chromosome alignment, and progression past the spindle assembly checkpoint in oocytes [PMID:35398309], and it associates with the mitotic spindle in neural progenitors where a truncating mutation stalls prometaphase and triggers p53-dependent apoptosis [PMID:41725360]. In differentiating epithelia, MAPRE2 drives initial microtubule reorganization by limiting EB1/ACF7 lattice association and formin-dependent actin co-alignment [PMID:23813963]. It promotes focal adhesion turnover and cell motility through a complex with HAX1 [PMID:26527684] and facilitates anterograde trafficking of the TRPM4 channel to the plasma membrane via an EB-binding motif on the channel [PMID:31112396]. In cardiomyocytes, MAPRE2 sustains voltage-gated sodium channel function and adherens junction integrity through a tubulin-detyrosination pathway, established by epistasis with the tubulin tyrosine ligase ttl [PMID:38095085]. MAPRE2 protein levels are set by ubiquitin-mediated degradation through the E3 ligases WDHD1 and ASB14 [PMID:32426268, PMID:38319584]. Gain-of-function mutations that increase MAPRE2 microtubule binding and plus-end dwell time perturb cranial neural crest migration and craniofacial development, causing circumferential skin creases Kunze type (CSC-KT) syndrome [PMID:26637975, PMID:33654163].","teleology":[{"year":2013,"claim":"Established that MAPRE2 actively shapes microtubule architecture during a developmental program rather than simply tracking plus ends, by showing it is required for the initial microtubule reorganization of epithelial differentiation.","evidence":"siRNA knockdown with live-cell imaging, formin-inhibitor rescue, and in situ tissue validation in epithelial cells","pmids":["23813963"],"confidence":"High","gaps":["Molecular basis for how EB2 restrains EB1/ACF7 lattice association not resolved","Direct binding partners mediating the actin co-alignment not identified"]},{"year":2015,"claim":"Linked MAPRE2 to a human Mendelian disease and defined the disease allele as gain-of-function, showing CSC-KT mutations enhance microtubule binding and disrupt craniofacial patterning.","evidence":"Microtubule co-sedimentation assays with patient mutants and zebrafish branchial arch analysis","pmids":["26637975"],"confidence":"High","gaps":["Cellular pathway connecting increased plus-end dwell time to craniofacial defects not yet defined","Effect on downstream effectors not measured"]},{"year":2015,"claim":"Identified a direct effector mechanism for MAPRE2 in cell migration by showing an EB2-HAX1 complex drives focal adhesion turnover.","evidence":"Quantitative proteomics with Co-IP validation, siRNA loss-of-function, in vitro migration and in vivo wound-healing assays","pmids":["26527684"],"confidence":"High","gaps":["Structural basis of the EB2-HAX1 interaction not determined","How the complex acts on focal adhesions mechanistically not resolved"]},{"year":2016,"claim":"Defined the regulatory logic of MAPRE2 in mitosis, showing Aurora B/CDK1 phosphorylation tunes its microtubule affinity to permit normal kinetochore microtubule dynamics and faithful chromosome segregation.","evidence":"In vitro kinase assays, non-phosphorylatable mutants, live-cell imaging and kinetochore microtubule dynamics assays","pmids":["27030108"],"confidence":"High","gaps":["Relative contributions of individual phospho-sites not dissected","Phosphatase that reverses these modifications not identified"]},{"year":2019,"claim":"Extended MAPRE2 function to membrane protein delivery by showing it is required for TRPM4 anterograde trafficking through an EB-binding motif.","evidence":"Co-IP with motif mutagenesis, immunofluorescence, cell invasion and focal adhesion assays","pmids":["31112396"],"confidence":"Medium","gaps":["EB1 and EB2 specific contributions not fully separated","Whether trafficking effect is direct or via microtubule dynamics unclear"]},{"year":2020,"claim":"Began to define how MAPRE2 abundance is controlled, identifying WDHD1 as an E3 ligase that ubiquitinates and degrades nuclear MAPRE2.","evidence":"Co-IP, ubiquitination and protein-stability assays, knockout/knockdown in cells and animals","pmids":["32426268"],"confidence":"Medium","gaps":["Single lab; reciprocal validation limited","Functional consequence of nuclear MAPRE2 pool not defined"]},{"year":2021,"claim":"Connected the CSC-KT gain-of-function allele to a cellular disease mechanism by showing mutant MAPRE2 alters neural crest focal adhesion dynamics and migration.","evidence":"Patient iPSC lines with isogenic CRISPR controls, neural crest differentiation, migration assays, chick xenotransplantation","pmids":["33654163"],"confidence":"Medium","gaps":["Link between altered plus-end dwell time and focal adhesion changes not mechanistically resolved","Single lab"]},{"year":2022,"claim":"Showed MAPRE2 spindle function is conserved in meiosis, being required for kinetochore-microtubule attachment and checkpoint satisfaction in oocytes.","evidence":"siRNA knockdown with spindle assembly checkpoint markers, cyclin B1 western blot and polar body extrusion assays in mouse oocytes","pmids":["35398309"],"confidence":"Medium","gaps":["Whether meiotic role is regulated by the same Aurora B/CDK1 phosphorylation not tested","Single lab"]},{"year":2023,"claim":"Defined a tissue-specific mechanism in heart, establishing by epistasis that MAPRE2 supports sodium channel function and adherens junctions through tubulin detyrosination.","evidence":"CRISPR zebrafish knockout and iPSC-CM knockdown with patch clamping, voltage mapping, plus-end tracking, and ttl-knockdown genetic rescue","pmids":["38095085"],"confidence":"High","gaps":["Direct link between MAPRE2 and the detyrosination enzymes not established","How detyrosinated tubulin regulates the channel not resolved"]},{"year":2024,"claim":"Added a second E3 ligase, ASB14, controlling MAPRE2 turnover, with degradation gating cardiomyocyte proliferation and cardiac repair.","evidence":"Microarray, ASB14 silencing in vitro and in vivo, western blot for MAPRE2, cardiomyocyte proliferation assays","pmids":["38319584"],"confidence":"Medium","gaps":["Mechanistic detail of ubiquitination limited","Relationship between ASB14- and WDHD1-mediated degradation not clarified"]},{"year":2024,"claim":"Positioned MAPRE2 as a phosphorylation substrate of CDKL5 in neurons, linking its modification to CDKL5 phase separation and disease variants.","evidence":"Optogenetic LLPS induction, super-resolution imaging, phosphorylation assays with CDKL5 truncation mutants, and patient CDD neurons (preprint)","pmids":["bio_10.1101_2024.11.18.624084","bio_10.1101_2024.11.11.622878"],"confidence":"Medium","gaps":["Functional consequence of CDKL5-mediated EB2 phosphorylation not defined","Preprint, not peer reviewed; single lab"]},{"year":2026,"claim":"Demonstrated that a disease truncation disrupts MAPRE2 spindle function in neural progenitors, stalling mitosis and triggering p53-dependent apoptosis as a likely basis for neurodevelopmental phenotypes.","evidence":"Transgenic mouse models and human stem cell-derived neural progenitors with time-lapse imaging and p53 pathway analysis","pmids":["41725360"],"confidence":"High","gaps":["Whether truncation acts as loss- or gain-of-function on the spindle not fully resolved","Link to focal-adhesion phenotypes of other mutants not established"]},{"year":null,"claim":"How MAPRE2's distinct cellular roles—spindle dynamics, focal adhesion turnover, membrane trafficking, and detyrosination signaling—are coordinated and differentially regulated across tissues remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unifying model linking phospho-regulation to tissue-specific outputs","Structural basis of mutant gain-of-function plus-end dwell time not determined","Direct enzymatic link to tubulin detyrosination not established"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[0,1,2]}],"localization":[{"term_id":"GO:0005815","term_label":"microtubule organizing center","supporting_discovery_ids":[0,9,13]},{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[0,1,8]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[5]}],"pathway":[{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[0,9,13]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[1,2,7]}],"complexes":[],"partners":["HAX1","TRPM4","WDHD1","ASB14","CDKL5","AURKB","CDK1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q15555","full_name":"Microtubule-associated protein RP/EB family member 2","aliases":["APC-binding protein EB2","End-binding protein 2","EB2"],"length_aa":327,"mass_kda":37.0,"function":"Adapter protein that is involved in microtubule polymerization, and spindle function by stabilizing microtubules and anchoring them at centrosomes. Therefore, ensures mitotic progression and genome stability (PubMed:27030108). Acts as a central regulator of microtubule reorganization in apico-basal epithelial differentiation (By similarity). Plays a role during oocyte meiosis by regulating microtubule dynamics (By similarity). Participates in neurite growth by interacting with plexin B3/PLXNB3 and microtubule reorganization during apico-basal epithelial differentiation (PubMed:22373814). Also plays an essential role for cell migration and focal adhesion dynamics. Mechanistically, recruits HAX1 to microtubules in order to regulate focal adhesion dynamics (PubMed:26527684)","subcellular_location":"Cytoplasm, cytoskeleton","url":"https://www.uniprot.org/uniprotkb/Q15555/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/MAPRE2","classification":"Not Classified","n_dependent_lines":1,"n_total_lines":1208,"dependency_fraction":0.0008278145695364238},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"CAPZB","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/MAPRE2","total_profiled":1310},"omim":[{"mim_id":"616734","title":"SKIN CREASES, CONGENITAL SYMMETRIC CIRCUMFERENTIAL, 2; CSCSC2","url":"https://www.omim.org/entry/616734"},{"mim_id":"605789","title":"MICROTUBULE-ASSOCIATED PROTEIN, RP/EB FAMILY, MEMBER 2; MAPRE2","url":"https://www.omim.org/entry/605789"},{"mim_id":"605788","title":"MICROTUBULE-ASSOCIATED PROTEIN, RP/EB FAMILY, MEMBER 3; MAPRE3","url":"https://www.omim.org/entry/605788"},{"mim_id":"604754","title":"ZINC FINGER PROTEIN 271, PSEUDOGENE; ZNF271P","url":"https://www.omim.org/entry/604754"},{"mim_id":"603108","title":"MICROTUBULE-ASSOCIATED PROTEIN, RP/EB FAMILY, MEMBER 1; MAPRE1","url":"https://www.omim.org/entry/603108"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"brain","ntpm":130.8}],"url":"https://www.proteinatlas.org/search/MAPRE2"},"hgnc":{"alias_symbol":["RP1","EB1","EB2"],"prev_symbol":[]},"alphafold":{"accession":"Q15555","domains":[{"cath_id":"1.10.418.10","chopping":"59-172","consensus_level":"high","plddt":95.1887,"start":59,"end":172},{"cath_id":"1.20.5.1430","chopping":"240-297","consensus_level":"high","plddt":88.6502,"start":240,"end":297}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q15555","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q15555-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q15555-F1-predicted_aligned_error_v6.png","plddt_mean":74.19},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=MAPRE2","jax_strain_url":"https://www.jax.org/strain/search?query=MAPRE2"},"sequence":{"accession":"Q15555","fasta_url":"https://rest.uniprot.org/uniprotkb/Q15555.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q15555/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q15555"}},"corpus_meta":[{"pmid":"3028777","id":"PMC_3028777","title":"Both Epstein-Barr virus (EBV)-encoded trans-acting factors, EB1 and EB2, are required to activate transcription from an EBV early promoter.","date":"1986","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/3028777","citation_count":358,"is_preprint":false},{"pmid":"2555554","id":"PMC_2555554","title":"The Epstein-Barr virus (EBV) early protein EB2 is a posttranscriptional activator expressed under the control of EBV transcription factors EB1 and R.","date":"1989","source":"Journal of virology","url":"https://pubmed.ncbi.nlm.nih.gov/2555554","citation_count":102,"is_preprint":false},{"pmid":"16129689","id":"PMC_16129689","title":"Interaction of the Epstein-Barr virus mRNA export factor EB2 with human Spen proteins SHARP, OTT1, and a novel member of the family, OTT3, links Spen proteins with splicing regulation and mRNA export.","date":"2005","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/16129689","citation_count":82,"is_preprint":false},{"pmid":"12403791","id":"PMC_12403791","title":"A novel nuclear export signal and a REF interaction domain both promote mRNA export by the Epstein-Barr virus EB2 protein.","date":"2002","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/12403791","citation_count":74,"is_preprint":false},{"pmid":"12208942","id":"PMC_12208942","title":"Epstein-Barr virus mRNA export factor EB2 is essential for production of infectious virus.","date":"2002","source":"Journal of virology","url":"https://pubmed.ncbi.nlm.nih.gov/12208942","citation_count":72,"is_preprint":false},{"pmid":"26637975","id":"PMC_26637975","title":"Mutations in Either TUBB or MAPRE2 Cause Circumferential Skin Creases Kunze Type.","date":"2015","source":"American journal of human 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This phosphorylation reduces MAPRE2 binding affinity for microtubules. Expression of non-phosphorylatable MAPRE2 stabilizes kinetochore microtubule dynamics, delays bipolar metaphase plate formation, and leads to aneuploidy.\",\n      \"method\": \"In vitro kinase assays, phospho-site mutagenesis (non-phosphorylatable mutants), live-cell imaging, kinetochore microtubule dynamics assays\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro kinase assay with mutagenesis plus defined cellular phenotype in single rigorous study\",\n      \"pmids\": [\"27030108\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"MAPRE2/EB2 is required for initial microtubule reorganisation during apico-basal epithelial differentiation. siRNA knockdown of EB2 in undifferentiated epithelial cells induced formation of straight, less dynamic microtubules with EB1 and ACF7 lattice association and co-alignment with actin filaments, a phenotype rescued by formin inhibition. Conversely, EB2 downregulation at later differentiation stages promoted microtubule stability and bundle formation.\",\n      \"method\": \"siRNA knockdown, live-cell imaging, immunofluorescence, formin inhibitor rescue experiments, in situ tissue analysis\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — siRNA knockdown with defined cellular phenotype, rescue experiment, and orthogonal in situ tissue validation\",\n      \"pmids\": [\"23813963\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"MAPRE2 mutations associated with circumferential skin creases Kunze type (CSC-KT) syndrome result in enhanced MAPRE2 binding to microtubules, implying an increased dwell time at microtubule plus ends. In a zebrafish craniofacial development model, MAPRE2 mutations perturb branchial arch patterning.\",\n      \"method\": \"Microtubule co-sedimentation/binding assays with patient-derived mutants, zebrafish in vivo craniofacial analysis\",\n      \"journal\": \"American journal of human genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — direct biochemical binding assay with mutants plus in vivo zebrafish model, single study with two orthogonal methods\",\n      \"pmids\": [\"26637975\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"MAPRE2/EB2 interacts specifically with HAX1 (HCLS1-associated protein X-1), identified by quantitative proteomics. Knockdown of HAX1 or EB2 in skin epidermal cells stabilizes focal adhesions and impairs epidermal migration in vitro and in vivo. The EB2-HAX1 interaction is required for focal adhesion turnover and cell motility.\",\n      \"method\": \"Quantitative proteomics (mass spectrometry), Co-immunoprecipitation, siRNA knockdown, in vitro migration assays, in vivo wound healing model\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — MS-identified interaction validated by Co-IP, loss-of-function with defined phenotype in vitro and in vivo\",\n      \"pmids\": [\"26527684\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"MAPRE2/EB2 mediates microtubule destabilization, increases Src kinase activity, and thereby facilitates ERK signaling activation in hepatocellular carcinoma cells. ERK signaling in turn promotes EB2 expression, forming a positive feedback loop. ERK inhibitor U0126 blocked EB2-mediated proliferation and metastasis.\",\n      \"method\": \"Overexpression and siRNA knockdown, in vitro invasion assays, in vivo xenograft, kinase activity assays, ERK inhibitor treatment\",\n      \"journal\": \"Clinical science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — defined pathway placement with in vitro and in vivo data but single lab, mechanism of Src activation by EB2 not directly demonstrated\",\n      \"pmids\": [\"33755094\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"WDHD1 (a ubiquitin ligase) interacts with MAPRE2 in the nucleus and promotes MAPRE2 ubiquitination, leading to MAPRE2 degradation and contributing to cisplatin resistance in lung adenocarcinoma cells.\",\n      \"method\": \"Co-immunoprecipitation, protein stability assays, ubiquitination assays, immunofluorescence, MAPRE2 and WDHD1 knockout/knockdown in cell lines and animal experiments\",\n      \"journal\": \"Frontiers in oncology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — Co-IP and ubiquitination assays with functional validation, single lab\",\n      \"pmids\": [\"32426268\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"E3 ubiquitin ligase ASB14 promotes MAPRE2 ubiquitination and degradation. Loss of ASB14 decreases MAPRE2 protein degradation, promotes cardiomyocyte nuclear proliferation, and enhances cardiac repair after myocardial infarction.\",\n      \"method\": \"Microarray analysis, ASB14 silencing in vitro and in vivo, Western blot for MAPRE2 protein levels, cardiomyocyte proliferation assays\",\n      \"journal\": \"Cell biochemistry and biophysics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — ubiquitination pathway placement with in vitro and in vivo data, single lab, mechanistic detail limited in abstract\",\n      \"pmids\": [\"38319584\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"MAPRE2 mutations cause altered cranial neural crest cell migration in vitro and in vivo (xenotransplantation into chicken embryos). Changes in focal adhesion dynamics underlie the altered cell motility of MAPRE2 mutant cranial neural crest cells.\",\n      \"method\": \"Patient-derived iPSC lines with CRISPR/Cas9 isogenic controls, neural crest differentiation, in vitro migration assays, xenotransplantation into chicken embryos, focal adhesion analysis\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — iPSC-derived disease models with isogenic controls, in vitro and in vivo validation, orthogonal methods\",\n      \"pmids\": [\"33654163\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"MAPRE2 loss-of-function (CRISPR knockout in zebrafish and siRNA/KO in human iPSC-CMs) decreases voltage-gated sodium channel function (reduced sodium current density) and disrupts adherens junctions with mislocalization of mature N-cadherin, associated with decreased detyrosinated tubulin. MAPRE2 knockdown in iPSC-CMs increases microtubule growth velocity and distance. Knockdown of ttl (tubulin tyrosine ligase) in mapre2 knockout larvae rescued detyrosinated tubulin levels and ventricular action potential upstroke velocity, establishing epistasis between MAPRE2 and tubulin detyrosination in regulating sodium channel function.\",\n      \"method\": \"CRISPR/Cas9 zebrafish knockout, siRNA knockdown and KO in iPSC-CMs, patch clamping, ECG, voltage mapping, immunocytochemistry, microtubule plus-end tracking, genetic epistasis (ttl knockdown rescue)\",\n      \"journal\": \"Circulation research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (electrophysiology, imaging, genetic epistasis rescue), in vivo and in vitro, with mechanistic pathway placement\",\n      \"pmids\": [\"38095085\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"MAPRE2 is distributed along the meiotic spindle during metaphase I and II in mouse oocytes. siRNA-mediated knockdown of Mapre2 severely impairs microtubule stability, kinetochore-microtubule attachment, and chromosome alignment, activates the spindle assembly checkpoint, prevents cyclin B1 degradation, and leads to failure of chromosome segregation and first polar body extrusion.\",\n      \"method\": \"siRNA knockdown, immunofluorescence, spindle assembly checkpoint markers, cyclin B1 western blot, polar body extrusion assay\",\n      \"journal\": \"Experimental cell research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — siRNA knockdown with multiple defined cellular readouts, single lab\",\n      \"pmids\": [\"35398309\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"MAPRE2/EB2 interacts with TRPM4 via a specific EB-binding motif on TRPM4. Mutations of this motif abolish the TRPM4-EB interaction and reduce plasma membrane expression of the channel. EB1 and EB2 proteins are required for TRPM4 anterograde trafficking; their loss leads to endoplasmic reticulum-associated TRPM4 distribution and affects focal adhesion disassembly and cell invasion.\",\n      \"method\": \"Co-immunoprecipitation, mutagenesis of EB-binding motif, immunofluorescence, cell invasion assays, focal adhesion assays\",\n      \"journal\": \"FASEB journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — Co-IP with mutagenesis and functional consequences, but EB1 and EB2 contributions not fully separated, single lab\",\n      \"pmids\": [\"31112396\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"MAPRE2 expression is significantly increased in highly nerve-invasive pancreatic cancer cells compared to less invasive cells, and changes in MAPRE2 expression result in altered actin distribution in these cells.\",\n      \"method\": \"siRNA knockdown/overexpression, immunofluorescence for actin distribution, comparative expression analysis\",\n      \"journal\": \"International journal of oncology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single method (actin redistribution by immunofluorescence) with no defined molecular mechanism, single lab\",\n      \"pmids\": [\"19787265\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Transcription factor JUND binds to the MAPRE2 promoter (shown by dual-luciferase reporter and ChIP assay) and increases MAPRE2 transcription, thereby facilitating cell proliferative and angiogenic abilities in esophageal squamous cell carcinoma.\",\n      \"method\": \"Dual-luciferase reporter assay, chromatin immunoprecipitation (ChIP), siRNA knockdown, in vitro angiogenesis and proliferation assays\",\n      \"journal\": \"Tissue & cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — direct promoter binding demonstrated by ChIP and reporter assay, functional rescue, single lab\",\n      \"pmids\": [\"36608637\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"EB2/MAPRE2 associates with the mitotic spindle in neural progenitors. A disease-causing truncation mutation (Q152X) stalls mitosis in prometaphase, causes defects in chromosome congression, misaligned chromosomes, and p53-induced apoptosis in both mouse transgenic models and human stem cell-derived neural progenitor cultures.\",\n      \"method\": \"Transgenic mouse models, stem cell-derived neural progenitor cultures, immunofluorescence, time-lapse imaging, p53 pathway analysis\",\n      \"journal\": \"Development (Cambridge, England)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — disease mutation in two orthogonal systems (mouse transgenics and human iPSC-derived cultures) with defined mitotic phenotype\",\n      \"pmids\": [\"41725360\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"CDKL5 phosphorylates MAPRE2/EB2, and this phosphorylation is impaired when CDKL5 undergoes truncating mutations (S726X, R781X) that disrupt its liquid-liquid phase separation. Reduced CDKL5 LLPS correlates with decreased EB2 phosphorylation.\",\n      \"method\": \"Optogenetic LLPS induction, super-resolution imaging, electron microscopy, phosphorylation assays with truncation mutants\",\n      \"journal\": \"bioRxiv (preprint)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — phosphorylation shown in cellular assay with disease mutants, preprint not yet peer reviewed, single lab\",\n      \"pmids\": [\"bio_10.1101_2024.11.18.624084\"],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Patient-derived CDD neurons (with loss-of-function CDKL5 variants) show decreased phosphorylated EB2, confirming EB2 as a molecular substrate of CDKL5 in human neurons.\",\n      \"method\": \"iPSC differentiation (NGN2-induced neurons and cortical organoids), Western blot for phospho-EB2\",\n      \"journal\": \"bioRxiv (preprint)\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — indirect evidence (reduced substrate phosphorylation in disease context), preprint, single readout\",\n      \"pmids\": [\"bio_10.1101_2024.11.11.622878\"],\n      \"is_preprint\": true\n    }\n  ],\n  \"current_model\": \"MAPRE2/EB2 is a microtubule plus-end tracking protein whose binding to microtubules is regulated by Aurora B- and CDK1-mediated phosphorylation during mitosis (and by CDKL5 phosphorylation in neurons), which controls kinetochore microtubule dynamics, mitotic progression, and genome stability; it promotes apico-basal epithelial differentiation by modulating microtubule reorganization and EB1/ACF7 deployment; it drives focal adhesion turnover and cell migration via an EB2–HAX1 complex; it facilitates TRPM4 anterograde trafficking to the plasma membrane; it regulates ventricular sodium channel function through a microtubule detyrosination pathway; its ubiquitination and degradation is mediated by the E3 ligases WDHD1 and ASB14; and gain-of-function mutations increase its dwell time at microtubule plus ends, perturbing neural crest migration and craniofacial development underlying CSC-KT syndrome.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"MAPRE2 (EB2) is a microtubule plus-end tracking protein that controls microtubule dynamics across mitosis, epithelial differentiation, cell migration, and membrane protein trafficking [#0, #1, #3]. During mitotic entry and progression, Aurora B and CDK1 phosphorylate MAPRE2 at the amino terminus and the linker between its calponin-homology and end-binding domains, reducing its microtubule binding affinity; blocking this phosphorylation stabilizes kinetochore microtubule dynamics, delays metaphase plate formation, and produces aneuploidy [#0]. Consistent with a conserved role in spindle function, MAPRE2 decorates the meiotic spindle and is required for kinetochore-microtubule attachment, chromosome alignment, and progression past the spindle assembly checkpoint in oocytes [#9], and it associates with the mitotic spindle in neural progenitors where a truncating mutation stalls prometaphase and triggers p53-dependent apoptosis [#13]. In differentiating epithelia, MAPRE2 drives initial microtubule reorganization by limiting EB1/ACF7 lattice association and formin-dependent actin co-alignment [#1]. It promotes focal adhesion turnover and cell motility through a complex with HAX1 [#3] and facilitates anterograde trafficking of the TRPM4 channel to the plasma membrane via an EB-binding motif on the channel [#10]. In cardiomyocytes, MAPRE2 sustains voltage-gated sodium channel function and adherens junction integrity through a tubulin-detyrosination pathway, established by epistasis with the tubulin tyrosine ligase ttl [#8]. MAPRE2 protein levels are set by ubiquitin-mediated degradation through the E3 ligases WDHD1 and ASB14 [#5, #6]. Gain-of-function mutations that increase MAPRE2 microtubule binding and plus-end dwell time perturb cranial neural crest migration and craniofacial development, causing circumferential skin creases Kunze type (CSC-KT) syndrome [#2, #7].\",\n  \"teleology\": [\n    {\n      \"year\": 2013,\n      \"claim\": \"Established that MAPRE2 actively shapes microtubule architecture during a developmental program rather than simply tracking plus ends, by showing it is required for the initial microtubule reorganization of epithelial differentiation.\",\n      \"evidence\": \"siRNA knockdown with live-cell imaging, formin-inhibitor rescue, and in situ tissue validation in epithelial cells\",\n      \"pmids\": [\"23813963\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular basis for how EB2 restrains EB1/ACF7 lattice association not resolved\", \"Direct binding partners mediating the actin co-alignment not identified\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Linked MAPRE2 to a human Mendelian disease and defined the disease allele as gain-of-function, showing CSC-KT mutations enhance microtubule binding and disrupt craniofacial patterning.\",\n      \"evidence\": \"Microtubule co-sedimentation assays with patient mutants and zebrafish branchial arch analysis\",\n      \"pmids\": [\"26637975\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Cellular pathway connecting increased plus-end dwell time to craniofacial defects not yet defined\", \"Effect on downstream effectors not measured\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Identified a direct effector mechanism for MAPRE2 in cell migration by showing an EB2-HAX1 complex drives focal adhesion turnover.\",\n      \"evidence\": \"Quantitative proteomics with Co-IP validation, siRNA loss-of-function, in vitro migration and in vivo wound-healing assays\",\n      \"pmids\": [\"26527684\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of the EB2-HAX1 interaction not determined\", \"How the complex acts on focal adhesions mechanistically not resolved\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Defined the regulatory logic of MAPRE2 in mitosis, showing Aurora B/CDK1 phosphorylation tunes its microtubule affinity to permit normal kinetochore microtubule dynamics and faithful chromosome segregation.\",\n      \"evidence\": \"In vitro kinase assays, non-phosphorylatable mutants, live-cell imaging and kinetochore microtubule dynamics assays\",\n      \"pmids\": [\"27030108\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Relative contributions of individual phospho-sites not dissected\", \"Phosphatase that reverses these modifications not identified\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Extended MAPRE2 function to membrane protein delivery by showing it is required for TRPM4 anterograde trafficking through an EB-binding motif.\",\n      \"evidence\": \"Co-IP with motif mutagenesis, immunofluorescence, cell invasion and focal adhesion assays\",\n      \"pmids\": [\"31112396\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"EB1 and EB2 specific contributions not fully separated\", \"Whether trafficking effect is direct or via microtubule dynamics unclear\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Began to define how MAPRE2 abundance is controlled, identifying WDHD1 as an E3 ligase that ubiquitinates and degrades nuclear MAPRE2.\",\n      \"evidence\": \"Co-IP, ubiquitination and protein-stability assays, knockout/knockdown in cells and animals\",\n      \"pmids\": [\"32426268\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab; reciprocal validation limited\", \"Functional consequence of nuclear MAPRE2 pool not defined\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Connected the CSC-KT gain-of-function allele to a cellular disease mechanism by showing mutant MAPRE2 alters neural crest focal adhesion dynamics and migration.\",\n      \"evidence\": \"Patient iPSC lines with isogenic CRISPR controls, neural crest differentiation, migration assays, chick xenotransplantation\",\n      \"pmids\": [\"33654163\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Link between altered plus-end dwell time and focal adhesion changes not mechanistically resolved\", \"Single lab\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Showed MAPRE2 spindle function is conserved in meiosis, being required for kinetochore-microtubule attachment and checkpoint satisfaction in oocytes.\",\n      \"evidence\": \"siRNA knockdown with spindle assembly checkpoint markers, cyclin B1 western blot and polar body extrusion assays in mouse oocytes\",\n      \"pmids\": [\"35398309\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether meiotic role is regulated by the same Aurora B/CDK1 phosphorylation not tested\", \"Single lab\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Defined a tissue-specific mechanism in heart, establishing by epistasis that MAPRE2 supports sodium channel function and adherens junctions through tubulin detyrosination.\",\n      \"evidence\": \"CRISPR zebrafish knockout and iPSC-CM knockdown with patch clamping, voltage mapping, plus-end tracking, and ttl-knockdown genetic rescue\",\n      \"pmids\": [\"38095085\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct link between MAPRE2 and the detyrosination enzymes not established\", \"How detyrosinated tubulin regulates the channel not resolved\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Added a second E3 ligase, ASB14, controlling MAPRE2 turnover, with degradation gating cardiomyocyte proliferation and cardiac repair.\",\n      \"evidence\": \"Microarray, ASB14 silencing in vitro and in vivo, western blot for MAPRE2, cardiomyocyte proliferation assays\",\n      \"pmids\": [\"38319584\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanistic detail of ubiquitination limited\", \"Relationship between ASB14- and WDHD1-mediated degradation not clarified\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Positioned MAPRE2 as a phosphorylation substrate of CDKL5 in neurons, linking its modification to CDKL5 phase separation and disease variants.\",\n      \"evidence\": \"Optogenetic LLPS induction, super-resolution imaging, phosphorylation assays with CDKL5 truncation mutants, and patient CDD neurons (preprint)\",\n      \"pmids\": [\"bio_10.1101_2024.11.18.624084\", \"bio_10.1101_2024.11.11.622878\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional consequence of CDKL5-mediated EB2 phosphorylation not defined\", \"Preprint, not peer reviewed; single lab\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Demonstrated that a disease truncation disrupts MAPRE2 spindle function in neural progenitors, stalling mitosis and triggering p53-dependent apoptosis as a likely basis for neurodevelopmental phenotypes.\",\n      \"evidence\": \"Transgenic mouse models and human stem cell-derived neural progenitors with time-lapse imaging and p53 pathway analysis\",\n      \"pmids\": [\"41725360\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether truncation acts as loss- or gain-of-function on the spindle not fully resolved\", \"Link to focal-adhesion phenotypes of other mutants not established\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How MAPRE2's distinct cellular roles—spindle dynamics, focal adhesion turnover, membrane trafficking, and detyrosination signaling—are coordinated and differentially regulated across tissues remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No unifying model linking phospho-regulation to tissue-specific outputs\", \"Structural basis of mutant gain-of-function plus-end dwell time not determined\", \"Direct enzymatic link to tubulin detyrosination not established\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [0, 1, 2]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005815\", \"supporting_discovery_ids\": [0, 9, 13]},\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [0, 1, 8]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [5]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [0, 9, 13]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [1, 2, 7]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"HAX1\", \"TRPM4\", \"WDHD1\", \"ASB14\", \"CDKL5\", \"AURKB\", \"CDK1\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":8,"faith_total":8,"faith_pct":100.0}}