{"gene":"MAP7D1","run_date":"2026-04-28T18:30:27","timeline":{"discoveries":[{"year":2016,"finding":"DCLK1 phosphorylates MAP7D1 at serine 315 to promote axon elongation in cortical neurons. Knockdown of MAP7D1 in layer 2/3 cortical neurons impairs callosal axon elongation (but not radial migration), and a phosphomimetic MAP7D1 S315E mutant fully rescues axon elongation defects in Dclk1 knockdown neurons.","method":"Proteomic substrate identification, in vitro kinase assay, in utero electroporation knockdown, phosphomimetic rescue experiment","journal":"Developmental neurobiology","confidence":"High","confidence_rationale":"Tier 2 — phosphorylation site identified biochemically, confirmed by phosphomimetic rescue in primary neurons with defined phenotypic readout","pmids":["27503845"],"is_preprint":false},{"year":2018,"finding":"MAP7D1 (together with MAP7) binds Disheveled, directs its cortical localization, and facilitates cortical targeting of microtubule plus-ends in response to Wnt5a signaling. Wnt5a signaling reciprocally promotes MAP7D1 movement toward microtubule plus-ends, and this dynamics requires Kinesin-1 member KIF5B. Disheveled also stabilizes MAP7D1. This MAP7/7D1–Disheveled feedback loop is evolutionarily conserved (Drosophila Ensconsin influences Disheveled localization in pupal wing cells).","method":"Co-immunoprecipitation, depletion/rescue experiments in HeLa cells, live-cell imaging, Drosophila genetic analysis","journal":"EMBO reports","confidence":"High","confidence_rationale":"Tier 2 — reciprocal Co-IP, functional depletion phenotypes, cross-species validation with ortholog","pmids":["29880710"],"is_preprint":false},{"year":2019,"finding":"All four mammalian MAP7 family members (MAP7, MAP7D1, MAP7D3, MAP7D2) bind to kinesin-1. MAP7, MAP7D1, and MAP7D3 act redundantly in HeLa cells to enable kinesin-1-dependent transport and microtubule recruitment of KIF5B-560. MAP7 proteins promote kinesin-1 binding to microtubules both directly (via the N-terminal microtubule-binding domain and unstructured linker) and indirectly via an allosteric effect from the kinesin-binding C-terminal domain.","method":"In vitro reconstitution with purified proteins, single-molecule TIRF assays, siRNA knockdown in HeLa cells, domain mapping","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 1 — in vitro reconstitution with purified proteins and single-molecule assays, complemented by cell-based knockdown","pmids":["30770434"],"is_preprint":false},{"year":2022,"finding":"MAP7D1 is required for maintenance of acetylated stable microtubules in neuronal cells. In contrast to Map7D2 (which stabilizes microtubules via direct binding), Map7D1 stabilizes microtubules through a distinct mechanism involving acetylation. Loss of Map7D1 increases random cell migration rate and neurite outgrowth.","method":"Gene knockdown (siRNA/shRNA), nocodazole resistance assay, immunofluorescence for acetylated/detyrosinated tubulin, live-cell migration assay in N1-E115 neuronal cells","journal":"Life science alliance","confidence":"Medium","confidence_rationale":"Tier 2 — clean KO/KD with defined cellular phenotypes and mechanistic distinction from paralog, single lab","pmids":["35470240"],"is_preprint":false},{"year":2023,"finding":"MAP7D1 interacts with DNA double-strand break repair proteins RAD50, BRCA1, and 53BP1. Downregulation of MAP7D1 causes strong G1 arrest and impairs DNA repair in G1-arrested cells, reducing RAD50 recruitment to chromatin and 53BP1 localization to damage sites, and increasing p53 phosphorylation after γ-irradiation.","method":"Quantitative proteomics (AP-MS), γ-irradiation, chromatin fractionation, immunofluorescence, siRNA knockdown","journal":"iScience","confidence":"Medium","confidence_rationale":"Tier 2 — MS-identified interactions confirmed by functional chromatin recruitment assays and defined cell cycle phenotype, single lab","pmids":["36852271"],"is_preprint":false},{"year":2021,"finding":"MAP7D1 disruption in zebrafish (map7d1b) exacerbates doxorubicin-induced cardiomyopathy; mechanistically, loss of MAP7D1 function impairs autophagic degradation and elevates protein aggregation. MAP7D1/Map7d1b shows cardiac and skeletal muscle-specific expression with sarcomeric localization.","method":"Zebrafish knockout, doxorubicin treatment, autophagic flux assays, protein aggregation assays, immunofluorescence","journal":"BioMed research international","confidence":"Medium","confidence_rationale":"Tier 2 — in vivo vertebrate model KO with defined mechanistic pathway (autophagy), single lab","pmids":["34327238"],"is_preprint":false},{"year":2021,"finding":"MAP7D1 expression is regulated by TET1-mediated 5-hydroxymethylcytosine modification and promotes tumor growth and metastasis in breast cancer.","method":"Genome-wide 5hmC profiling, TET1 knockdown/overexpression, functional metastasis assays","journal":"Genomics, proteomics & bioinformatics","confidence":"Medium","confidence_rationale":"Tier 2 — epigenetic regulation linked to functional metastasis phenotype, single lab","pmids":["33716151"],"is_preprint":false},{"year":2025,"finding":"A MAP7D1 R201W mutation in the microtubule-binding domain (found in a Shwachman-Diamond syndrome patient) acts as a loss-of-function mutation, disrupting MAP7D1 interaction with microtubules. This causes mitotic defects (multipolar/unstable spindles, lagging chromosomes, shortened inter-centrosomal distance) and accumulation of ribosomal protein RPS14 in dividing cells. Overexpression of mutant MAP7D1 or MAP7D1 depletion recapitulates these phenotypes in T98G and HEK293T cells.","method":"Patient fibroblast analysis, mutant overexpression, siRNA knockdown, immunofluorescence for spindle morphology and RPS14, microtubule co-sedimentation","journal":"Disease models & mechanisms","confidence":"Medium","confidence_rationale":"Tier 2 — patient variant with domain-specific mechanism validated in multiple cell lines, single lab","pmids":["40856631"],"is_preprint":false},{"year":2025,"finding":"MAP7D1 preferentially partitions onto detyrosinated microtubules (mediated by expanded microtubule lattice states), creating specialized tracks for kinesin-1 (KIF5B). MAP7D1 density on microtubules is dynamically modulated by nutrient availability — increasing during starvation (promoting perinuclear lysosome positioning) and decreasing upon nutrient stimulation (allowing peripheral lysosome migration). Altering MAP7D1 levels in either direction impairs lysosomal motility and nutrient-responsive positioning.","method":"Live-cell imaging, fluorescence microscopy, siRNA knockdown and overexpression, lysosome tracking, rigor kinesin localization assays","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 2 — multiple orthogonal imaging and functional methods, but preprint not yet peer-reviewed","pmids":["bio_10.1101_2025.10.07.680844"],"is_preprint":true},{"year":2020,"finding":"SARS-CoV-2 ORF10 protein physically associates with MAP7D1 in human cells, as identified by affinity-purification mass spectrometry.","method":"Affinity-purification mass spectrometry (AP-MS) in human cells expressing tagged viral proteins","journal":"Nature","confidence":"Low","confidence_rationale":"Tier 3 — single AP-MS identification in a large-scale screen; no follow-up mechanistic validation of MAP7D1 function","pmids":["32353859"],"is_preprint":false}],"current_model":"MAP7D1 is a microtubule-associated protein that preferentially binds detyrosinated microtubules and acts as a microtubule-tethered activator of kinesin-1, promoting motor landing and processivity; it is phosphorylated by DCLK1 at Ser315 to drive axon elongation, interacts with Disheveled to regulate Wnt5a-dependent cortical microtubule plus-end targeting, maintains acetylated stable microtubules in neuronal cells, participates in DNA double-strand break repair in G1 by recruiting RAD50 and 53BP1 to damage sites, controls lysosome positioning in response to nutrient signals, and its loss-of-function causes mitotic spindle defects and impaired autophagy."},"narrative":{"teleology":[{"year":2016,"claim":"Establishing MAP7D1 as a phosphorylation-dependent effector of axon elongation resolved how DCLK1 kinase activity is transduced into cytoskeletal remodeling during cortical neuron development.","evidence":"Proteomic substrate identification, in vitro kinase assay, and in utero electroporation with phosphomimetic rescue in cortical neurons","pmids":["27503845"],"confidence":"High","gaps":["Downstream mechanism by which Ser315 phosphorylation alters microtubule dynamics is unknown","Whether other kinases phosphorylate MAP7D1 at additional sites in neurons is untested"]},{"year":2018,"claim":"Demonstrating that MAP7D1 bridges Disheveled and kinesin-1 on microtubules established a Wnt5a-responsive feedback loop for cortical microtubule plus-end targeting, revealing a signaling-to-transport coupling mechanism.","evidence":"Reciprocal co-immunoprecipitation, live-cell imaging in HeLa cells, depletion/rescue, and cross-species validation in Drosophila","pmids":["29880710"],"confidence":"High","gaps":["Structural basis of the MAP7D1–Disheveled interaction is unresolved","Physiological tissue context beyond cultured cells and Drosophila wings is unexplored"]},{"year":2019,"claim":"Reconstitution of MAP7D1–kinesin-1 interactions with purified proteins demonstrated that MAP7 family members promote kinesin-1 microtubule landing through both direct tethering and allosteric activation, defining the core molecular mechanism of MAP7D1 as a motor cofactor.","evidence":"Single-molecule TIRF assays with purified proteins, domain truncation mapping, siRNA knockdown in HeLa cells","pmids":["30770434"],"confidence":"High","gaps":["Relative contributions of MAP7D1 versus MAP7 and MAP7D3 in specific tissues remain undefined","No high-resolution structure of the MAP7D1–kinesin-1 complex exists"]},{"year":2021,"claim":"Zebrafish map7d1b knockout revealed that MAP7D1 supports autophagic flux and protein homeostasis in cardiac muscle, extending its functional repertoire beyond cytoskeletal regulation to organelle quality control.","evidence":"Zebrafish knockout, doxorubicin-induced cardiomyopathy model, autophagic flux and protein aggregation assays","pmids":["34327238"],"confidence":"Medium","gaps":["Whether autophagy impairment is a direct consequence of disrupted kinesin-1 transport or an independent mechanism is unclear","Mammalian cardiac phenotypes of MAP7D1 loss have not been reported"]},{"year":2022,"claim":"Distinguishing MAP7D1's microtubule-stabilizing mechanism (via acetylation) from MAP7D2's direct binding-based stabilization clarified how paralog-specific functions diversify the microtubule cytoskeleton in neuronal cells.","evidence":"siRNA/shRNA knockdown, nocodazole resistance assay, acetylated/detyrosinated tubulin immunofluorescence in N1-E115 neuronal cells","pmids":["35470240"],"confidence":"Medium","gaps":["How MAP7D1 promotes tubulin acetylation (e.g., via αTAT1 recruitment) is unknown","In vivo neuronal consequences of selective MAP7D1 loss on stable microtubule pools are untested"]},{"year":2023,"claim":"Discovery that MAP7D1 recruits RAD50 and 53BP1 to DNA damage sites in G1 cells established an unexpected nuclear function for this microtubule-associated protein in the DNA double-strand break repair pathway.","evidence":"Quantitative AP-MS, γ-irradiation, chromatin fractionation, immunofluorescence, siRNA knockdown","pmids":["36852271"],"confidence":"Medium","gaps":["Whether MAP7D1 translocates to the nucleus or acts via an indirect cytoskeletal-signaling mechanism is unresolved","The DNA repair function has not been independently replicated"]},{"year":2025,"claim":"Identification of a patient-derived MAP7D1 R201W mutation that disrupts microtubule binding and causes multipolar spindles and lagging chromosomes provided the first direct link between MAP7D1 dysfunction and mitotic fidelity defects in a disease context.","evidence":"Patient fibroblast analysis, mutant overexpression, siRNA knockdown, microtubule co-sedimentation, spindle morphology analysis in T98G and HEK293T cells","pmids":["40856631"],"confidence":"Medium","gaps":["Whether MAP7D1 R201W is causative for the patient's Shwachman-Diamond syndrome phenotype or a modifier is not established","Mechanism linking MAP7D1 loss to RPS14 accumulation in dividing cells is unexplained"]},{"year":2025,"claim":"Showing that MAP7D1 preferentially partitions onto detyrosinated microtubules and that its density is nutrient-regulated to control lysosome positioning unified its kinesin-1 cofactor and tubulin-code reader activities into a nutrient-responsive transport mechanism.","evidence":"(preprint) Live-cell imaging, siRNA knockdown and overexpression, lysosome tracking, rigor kinesin localization assays","pmids":["bio_10.1101_2025.10.07.680844"],"confidence":"Medium","gaps":["Findings are from a preprint not yet peer-reviewed","Signaling pathway linking nutrient status to MAP7D1 density changes is unidentified","Whether detyrosination preference applies in vivo in specific tissues is untested"]},{"year":null,"claim":"A high-resolution structure of MAP7D1 bound to microtubules and/or kinesin-1, the molecular basis of its nuclear DNA repair role, and how nutrient signals regulate MAP7D1–microtubule partitioning remain major unresolved questions.","evidence":"","pmids":[],"confidence":"High","gaps":["No structural data for MAP7D1 or its complexes exist","Nuclear versus cytoplasmic partitioning mechanism is unknown","Tissue-specific functional redundancy among MAP7 family paralogs is poorly defined"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[0,1,2,3,7,8]}],"localization":[{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[0,1,2,3,7,8]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[4]}],"pathway":[{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[0,1]},{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[7]},{"term_id":"R-HSA-73894","term_label":"DNA Repair","supporting_discovery_ids":[4]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[1]}],"complexes":[],"partners":["KIF5B","DVL2","DCLK1","RAD50","TP53BP1","BRCA1"],"other_free_text":[]},"mechanistic_narrative":"MAP7D1 is a microtubule-associated protein that functions as a microtubule-tethered activator of kinesin-1 and a regulator of microtubule stability, with roles spanning axon elongation, mitotic spindle integrity, DNA damage repair, and organelle positioning. MAP7D1 binds microtubules through its N-terminal domain and promotes kinesin-1 (KIF5B) landing and processivity both directly and allosterically through its C-terminal kinesin-binding domain, acting redundantly with MAP7 and MAP7D3 in HeLa cells [PMID:30770434]. DCLK1 phosphorylates MAP7D1 at Ser315 to drive callosal axon elongation in cortical neurons [PMID:27503845], while MAP7D1 also interacts with Disheveled to mediate Wnt5a-dependent cortical microtubule plus-end targeting via a kinesin-1-dependent feedback loop [PMID:29880710]. Beyond cytoskeletal functions, MAP7D1 participates in G1-phase DNA double-strand break repair by recruiting RAD50 and 53BP1 to damage sites [PMID:36852271], maintains acetylated stable microtubules in neuronal cells [PMID:35470240], and its loss of function causes multipolar spindles and lagging chromosomes during mitosis [PMID:40856631]."},"prefetch_data":{"uniprot":{"accession":"Q3KQU3","full_name":"MAP7 domain-containing protein 1","aliases":["Arginine/proline-rich coiled-coil domain-containing protein 1","Proline/arginine-rich coiled-coil domain-containing protein 1"],"length_aa":841,"mass_kda":92.8,"function":"Microtubule-stabilizing protein involved in the control of cell motility and neurite outgrowth. Facilitate microtubule stabilization through the maintenance of acetylated stable microtubules","subcellular_location":"Cytoplasm, cytoskeleton, spindle; Cytoplasm, cytoskeleton; Cytoplasm, cytoskeleton, microtubule organizing center, centrosome; Midbody","url":"https://www.uniprot.org/uniprotkb/Q3KQU3/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/MAP7D1","classification":"Not 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MAP7D3","url":"https://www.omim.org/entry/300930"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"skeletal muscle","ntpm":267.9}],"url":"https://www.proteinatlas.org/search/MAP7D1"},"hgnc":{"alias_symbol":["FLJ10350","FLJ39022"],"prev_symbol":["PARCC1","RPRC1"]},"alphafold":{"accession":"Q3KQU3","domains":[{"cath_id":"-","chopping":"132-224","consensus_level":"high","plddt":88.8297,"start":132,"end":224},{"cath_id":"1.20.5","chopping":"619-694","consensus_level":"medium","plddt":88.5768,"start":619,"end":694}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q3KQU3","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q3KQU3-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q3KQU3-F1-predicted_aligned_error_v6.png","plddt_mean":58.44},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=MAP7D1","jax_strain_url":"https://www.jax.org/strain/search?query=MAP7D1"},"sequence":{"accession":"Q3KQU3","fasta_url":"https://rest.uniprot.org/uniprotkb/Q3KQU3.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q3KQU3/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q3KQU3"}},"corpus_meta":[{"pmid":"10744690","id":"PMC_10744690","title":"The 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biology","url":"https://pubmed.ncbi.nlm.nih.gov/27182664","citation_count":114,"is_preprint":false,"source_track":"gene2pubmed"}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":8287,"output_tokens":2117,"usd":0.028308},"stage2":{"model":"claude-opus-4-6","input_tokens":5373,"output_tokens":2415,"usd":0.13086},"total_usd":0.349129,"stage1_batch_id":"msgbatch_01Mgbec4vM6c9vXht7JVJzss","stage2_batch_id":"msgbatch_01JkuuH5RB2BdJNsgnhjbC6s","note":"batch pricing = 50% of standard","round2_stage1":{"model":"claude-sonnet-4-6","input_tokens":20026,"output_tokens":2621,"usd":0.049696},"round2_rules_fired":"R3","round2_stage2":{"model":"claude-opus-4-6","input_tokens":6182,"output_tokens":2504,"usd":0.140265}},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2016,\n      \"finding\": \"DCLK1 phosphorylates MAP7D1 at serine 315 (Ser315) to promote axon elongation in cortical neurons. Knockdown of MAP7D1 impairs callosal axon elongation, and a phosphomimetic MAP7D1 S315E mutant rescues axon elongation defects in Dclk1 knockdown neurons, demonstrating MAP7D1 is a direct substrate of DCLK1 in this pathway.\",\n      \"method\": \"Proteomic substrate identification, in vitro kinase assay, phosphosite mutagenesis (S315E phosphomimetic), in vivo knockdown/rescue in layer 2/3 cortical neurons\",\n      \"journal\": \"Developmental neurobiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — in vitro kinase assay with phosphosite mutagenesis plus in vivo rescue experiment; multiple orthogonal methods in single study\",\n      \"pmids\": [\"27503845\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"MAP7D1 (and its paralog Map7) binds to Disheveled (Dvl), directs its cortical localization, and facilitates cortical targeting of microtubule plus-ends in response to Wnt5a signaling. Kinesin-1 member Kif5b is required for Map7/7D1 dynamics and Disheveled localization. Conversely, Disheveled stabilizes Map7/7D1, forming a feedback regulatory loop. This role is evolutionarily conserved, as the Drosophila ortholog Ensconsin influences Drosophila Disheveled localization in pupal wing cells.\",\n      \"method\": \"Co-immunoprecipitation, siRNA knockdown, live-cell imaging, genetic epistasis in Drosophila, subcellular localization experiments in HeLa cells\",\n      \"journal\": \"EMBO reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal Co-IP, loss-of-function with defined cellular phenotypes, and evolutionary validation across two organisms\",\n      \"pmids\": [\"29880710\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Map7D1 facilitates microtubule stabilization in neuronal cells through a mechanism dependent on maintaining acetylated stable microtubules, distinct from Map7D2 which stabilizes microtubules via direct binding. Map7D1 loss decreases acetylated stable MTs without affecting nocodazole resistance, and Map7D1 knockdown alters cell motility and neurite outgrowth.\",\n      \"method\": \"Gene knockdown (siRNA/shRNA), immunofluorescence for acetylated/detyrosinated MTs, nocodazole resistance assay, live-cell migration assay in N1-E115 neuronal cells\",\n      \"journal\": \"Life science alliance\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — clean KO/KD with defined cellular phenotype and mechanistic distinction from paralog, single lab\",\n      \"pmids\": [\"35470240\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"MAP7D1 interacts with DNA double-strand break repair proteins RAD50, BRCA1, and 53BP1. Downregulation of MAP7D1 leads to a strong G1 cell cycle arrest, impairs DNA repair in G1-arrested cells, reduces RAD50 recruitment to chromatin, and disrupts 53BP1 localization to damage sites, revealing a novel role for MAP7D1 in DNA double-strand break repair.\",\n      \"method\": \"Quantitative proteomics (mass spectrometry), siRNA knockdown, γ-irradiation, chromatin fractionation, immunofluorescence for 53BP1 and RAD50 foci, flow cytometry for cell cycle analysis\",\n      \"journal\": \"iScience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — quantitative proteomics interaction data plus loss-of-function with defined repair phenotype, single lab\",\n      \"pmids\": [\"36852271\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"MAP7D1 (Map7d1b) localizes to the sarcomere in cardiac and skeletal muscle in zebrafish and mice. Disruption of map7d1b function in zebrafish exacerbates doxorubicin-induced cardiomyopathy, mechanistically conveyed through impaired autophagic degradation and elevated protein aggregation.\",\n      \"method\": \"Zebrafish genetic loss-of-function, cardiac phenotype assessment, autophagy flux assays, protein aggregation assays, expression validation in mice by immunofluorescence\",\n      \"journal\": \"BioMed research international\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — genetic loss-of-function in animal model with defined mechanistic pathway (autophagy impairment), single lab\",\n      \"pmids\": [\"34327238\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"A MAP7D1 R201W mutation in the microtubule-binding domain disrupts its interaction with microtubules, causing mitotic defects (multipolar/unstable spindles, lagging chromosomes, shortened inter-centrosomal distance) and accumulation of ribosomal protein RPS14. These phenotypes are recapitulated by overexpression of mutant MAP7D1 or depletion of MAP7D1 in cell lines, indicating the mutation acts as loss-of-function.\",\n      \"method\": \"Patient fibroblast analysis, microtubule co-sedimentation/binding assay, overexpression of mutant MAP7D1, siRNA depletion in T98G and HEK293T cells, immunofluorescence for spindle and RPS14 localization\",\n      \"journal\": \"Disease models & mechanisms\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — domain-disrupting mutation with functional validation in both patient cells and orthogonal cell lines, single lab\",\n      \"pmids\": [\"40856631\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"MAP7D1 selectively partitions onto detyrosinated microtubule subsets (while MAP4 partitions onto tyrosinated MTs), and this preferential binding is mediated by the expanded microtubule lattice state on detyrosinated MTs. MAP7D1-coated detyrosinated tracks preferentially recruit kinesin-1 (KIF5B). During nutrient starvation, MAP7D1 density on MTs increases and MAP4 density decreases, directing lysosomes to the perinuclear region; upon nutrient stimulation, MAP7D1 density declines and lysosomes migrate to the cell periphery. Altering MAP7D1 levels disrupts lysosomal motility and nutrient responsiveness.\",\n      \"method\": \"Live-cell imaging, immunofluorescence, rigor kinesin colocalization, MAP7D1 overexpression/knockdown, nutrient starvation/stimulation assays, lysosome positioning quantification\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods with functional consequence, but preprint without peer review\",\n      \"pmids\": [\"bio_10.1101_2025.10.07.680844\"],\n      \"is_preprint\": true\n    }\n  ],\n  \"current_model\": \"MAP7D1 is a microtubule-associated protein that stabilizes microtubules (particularly acetylated stable MT pools), is phosphorylated by DCLK1 at Ser315 to promote axonal elongation, binds Disheveled to couple microtubule plus-end dynamics to Wnt5a signaling, preferentially decorates detyrosinated microtubules to create kinesin-1 tracks that control lysosome positioning in nutrient signaling, participates in DNA double-strand break repair in G1 by facilitating RAD50 and 53BP1 recruitment, and plays roles in cardiac sarcomere integrity and mitotic spindle stability.\"\n}\n```\n\n--- ROUND 2 (R2/R3, augmented corpus) ---\n\n```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2016,\n      \"finding\": \"DCLK1 phosphorylates MAP7D1 at serine 315 to promote axon elongation in cortical neurons. Knockdown of MAP7D1 in layer 2/3 cortical neurons impairs callosal axon elongation (but not radial migration), and a phosphomimetic MAP7D1 S315E mutant fully rescues axon elongation defects in Dclk1 knockdown neurons.\",\n      \"method\": \"Proteomic substrate identification, in vitro kinase assay, in utero electroporation knockdown, phosphomimetic rescue experiment\",\n      \"journal\": \"Developmental neurobiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — phosphorylation site identified biochemically, confirmed by phosphomimetic rescue in primary neurons with defined phenotypic readout\",\n      \"pmids\": [\"27503845\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"MAP7D1 (together with MAP7) binds Disheveled, directs its cortical localization, and facilitates cortical targeting of microtubule plus-ends in response to Wnt5a signaling. Wnt5a signaling reciprocally promotes MAP7D1 movement toward microtubule plus-ends, and this dynamics requires Kinesin-1 member KIF5B. Disheveled also stabilizes MAP7D1. This MAP7/7D1–Disheveled feedback loop is evolutionarily conserved (Drosophila Ensconsin influences Disheveled localization in pupal wing cells).\",\n      \"method\": \"Co-immunoprecipitation, depletion/rescue experiments in HeLa cells, live-cell imaging, Drosophila genetic analysis\",\n      \"journal\": \"EMBO reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal Co-IP, functional depletion phenotypes, cross-species validation with ortholog\",\n      \"pmids\": [\"29880710\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"All four mammalian MAP7 family members (MAP7, MAP7D1, MAP7D3, MAP7D2) bind to kinesin-1. MAP7, MAP7D1, and MAP7D3 act redundantly in HeLa cells to enable kinesin-1-dependent transport and microtubule recruitment of KIF5B-560. MAP7 proteins promote kinesin-1 binding to microtubules both directly (via the N-terminal microtubule-binding domain and unstructured linker) and indirectly via an allosteric effect from the kinesin-binding C-terminal domain.\",\n      \"method\": \"In vitro reconstitution with purified proteins, single-molecule TIRF assays, siRNA knockdown in HeLa cells, domain mapping\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro reconstitution with purified proteins and single-molecule assays, complemented by cell-based knockdown\",\n      \"pmids\": [\"30770434\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"MAP7D1 is required for maintenance of acetylated stable microtubules in neuronal cells. In contrast to Map7D2 (which stabilizes microtubules via direct binding), Map7D1 stabilizes microtubules through a distinct mechanism involving acetylation. Loss of Map7D1 increases random cell migration rate and neurite outgrowth.\",\n      \"method\": \"Gene knockdown (siRNA/shRNA), nocodazole resistance assay, immunofluorescence for acetylated/detyrosinated tubulin, live-cell migration assay in N1-E115 neuronal cells\",\n      \"journal\": \"Life science alliance\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — clean KO/KD with defined cellular phenotypes and mechanistic distinction from paralog, single lab\",\n      \"pmids\": [\"35470240\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"MAP7D1 interacts with DNA double-strand break repair proteins RAD50, BRCA1, and 53BP1. Downregulation of MAP7D1 causes strong G1 arrest and impairs DNA repair in G1-arrested cells, reducing RAD50 recruitment to chromatin and 53BP1 localization to damage sites, and increasing p53 phosphorylation after γ-irradiation.\",\n      \"method\": \"Quantitative proteomics (AP-MS), γ-irradiation, chromatin fractionation, immunofluorescence, siRNA knockdown\",\n      \"journal\": \"iScience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — MS-identified interactions confirmed by functional chromatin recruitment assays and defined cell cycle phenotype, single lab\",\n      \"pmids\": [\"36852271\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"MAP7D1 disruption in zebrafish (map7d1b) exacerbates doxorubicin-induced cardiomyopathy; mechanistically, loss of MAP7D1 function impairs autophagic degradation and elevates protein aggregation. MAP7D1/Map7d1b shows cardiac and skeletal muscle-specific expression with sarcomeric localization.\",\n      \"method\": \"Zebrafish knockout, doxorubicin treatment, autophagic flux assays, protein aggregation assays, immunofluorescence\",\n      \"journal\": \"BioMed research international\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — in vivo vertebrate model KO with defined mechanistic pathway (autophagy), single lab\",\n      \"pmids\": [\"34327238\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"MAP7D1 expression is regulated by TET1-mediated 5-hydroxymethylcytosine modification and promotes tumor growth and metastasis in breast cancer.\",\n      \"method\": \"Genome-wide 5hmC profiling, TET1 knockdown/overexpression, functional metastasis assays\",\n      \"journal\": \"Genomics, proteomics & bioinformatics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — epigenetic regulation linked to functional metastasis phenotype, single lab\",\n      \"pmids\": [\"33716151\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"A MAP7D1 R201W mutation in the microtubule-binding domain (found in a Shwachman-Diamond syndrome patient) acts as a loss-of-function mutation, disrupting MAP7D1 interaction with microtubules. This causes mitotic defects (multipolar/unstable spindles, lagging chromosomes, shortened inter-centrosomal distance) and accumulation of ribosomal protein RPS14 in dividing cells. Overexpression of mutant MAP7D1 or MAP7D1 depletion recapitulates these phenotypes in T98G and HEK293T cells.\",\n      \"method\": \"Patient fibroblast analysis, mutant overexpression, siRNA knockdown, immunofluorescence for spindle morphology and RPS14, microtubule co-sedimentation\",\n      \"journal\": \"Disease models & mechanisms\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — patient variant with domain-specific mechanism validated in multiple cell lines, single lab\",\n      \"pmids\": [\"40856631\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"MAP7D1 preferentially partitions onto detyrosinated microtubules (mediated by expanded microtubule lattice states), creating specialized tracks for kinesin-1 (KIF5B). MAP7D1 density on microtubules is dynamically modulated by nutrient availability — increasing during starvation (promoting perinuclear lysosome positioning) and decreasing upon nutrient stimulation (allowing peripheral lysosome migration). Altering MAP7D1 levels in either direction impairs lysosomal motility and nutrient-responsive positioning.\",\n      \"method\": \"Live-cell imaging, fluorescence microscopy, siRNA knockdown and overexpression, lysosome tracking, rigor kinesin localization assays\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal imaging and functional methods, but preprint not yet peer-reviewed\",\n      \"pmids\": [\"bio_10.1101_2025.10.07.680844\"],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"SARS-CoV-2 ORF10 protein physically associates with MAP7D1 in human cells, as identified by affinity-purification mass spectrometry.\",\n      \"method\": \"Affinity-purification mass spectrometry (AP-MS) in human cells expressing tagged viral proteins\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — single AP-MS identification in a large-scale screen; no follow-up mechanistic validation of MAP7D1 function\",\n      \"pmids\": [\"32353859\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"MAP7D1 is a microtubule-associated protein that preferentially binds detyrosinated microtubules and acts as a microtubule-tethered activator of kinesin-1, promoting motor landing and processivity; it is phosphorylated by DCLK1 at Ser315 to drive axon elongation, interacts with Disheveled to regulate Wnt5a-dependent cortical microtubule plus-end targeting, maintains acetylated stable microtubules in neuronal cells, participates in DNA double-strand break repair in G1 by recruiting RAD50 and 53BP1 to damage sites, controls lysosome positioning in response to nutrient signals, and its loss-of-function causes mitotic spindle defects and impaired autophagy.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"MAP7D1 is a microtubule-associated protein that stabilizes acetylated microtubule pools, preferentially decorates detyrosinated microtubules to create kinesin-1 tracks, and participates in axonal elongation, Wnt signaling, DNA repair, and mitotic spindle integrity. In neurons, MAP7D1 is phosphorylated by DCLK1 at Ser315 to promote axon elongation, and it binds Disheveled to couple microtubule plus-end dynamics to Wnt5a-dependent cortical polarity signaling [PMID:27503845, PMID:29880710]. MAP7D1 maintains acetylated stable microtubule subsets distinct from the direct microtubule-stabilizing mechanism of its paralog MAP7D2, and loss of MAP7D1 impairs cell motility and neurite outgrowth [PMID:35470240]. Beyond cytoskeletal functions, MAP7D1 interacts with RAD50, BRCA1, and 53BP1 to facilitate DNA double-strand break repair in G1-phase cells, and disruption of its microtubule-binding domain causes mitotic spindle defects including multipolar spindles and lagging chromosomes [PMID:36852271, PMID:40856631].\",\n  \"teleology\": [\n    {\n      \"year\": 2016,\n      \"claim\": \"Identifying MAP7D1 as a direct DCLK1 substrate established that a specific phosphorylation event (Ser315) couples doublecortin-like kinase signaling to microtubule-dependent axon elongation.\",\n      \"evidence\": \"In vitro kinase assay, phosphomimetic S315E mutagenesis, and in vivo knockdown/rescue in layer 2/3 cortical neurons\",\n      \"pmids\": [\"27503845\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"How Ser315 phosphorylation alters MAP7D1 conformation or microtubule binding is unknown\",\n        \"Whether other kinases phosphorylate MAP7D1 at additional sites in vivo is untested\",\n        \"Relevance beyond cortical projection neurons has not been examined\"\n      ]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Demonstrating that MAP7D1 binds Disheveled, directs its cortical localization, and requires kinesin-1 (Kif5b) revealed a feedback loop linking MAP7D1–microtubule dynamics to Wnt5a planar cell polarity signaling.\",\n      \"evidence\": \"Reciprocal co-immunoprecipitation, siRNA knockdown, live-cell imaging in HeLa cells, and genetic epistasis with Drosophila Ensconsin in pupal wing cells\",\n      \"pmids\": [\"29880710\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"The direct binding interface between MAP7D1 and Disheveled has not been mapped\",\n        \"Whether this interaction operates in non-epithelial contexts such as neurons is unresolved\",\n        \"Downstream transcriptional consequences of disrupted Dvl cortical targeting are not characterized\"\n      ]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Localizing MAP7D1 to cardiac and skeletal muscle sarcomeres and showing that its loss exacerbates doxorubicin-induced cardiomyopathy through impaired autophagy extended its known roles beyond neurons to striated muscle homeostasis.\",\n      \"evidence\": \"Zebrafish genetic loss-of-function, autophagy flux and protein aggregation assays, immunofluorescence validation in mouse tissue\",\n      \"pmids\": [\"34327238\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"The mechanism by which MAP7D1 promotes autophagic degradation in cardiomyocytes is unclear\",\n        \"Sarcomeric binding partners have not been identified\",\n        \"Whether mammalian cardiac-specific knockout recapitulates the zebrafish phenotype is untested\"\n      ]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Distinguishing MAP7D1's stabilization of acetylated microtubule pools from MAP7D2's direct microtubule-binding stabilization mechanism clarified how paralogous MAP7 family members partition microtubule regulatory functions.\",\n      \"evidence\": \"siRNA/shRNA knockdown in N1-E115 neuronal cells, immunofluorescence for acetylated and detyrosinated tubulin marks, nocodazole resistance assay\",\n      \"pmids\": [\"35470240\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Whether MAP7D1 promotes acetylation indirectly (e.g., via αTAT1 recruitment) or by protecting acetylated MTs from depolymerization is unresolved\",\n        \"Functional redundancy with MAP7 in vivo has not been systematically tested\"\n      ]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Identifying MAP7D1 interactions with RAD50, BRCA1, and 53BP1 and showing that MAP7D1 depletion impairs G1-phase DNA double-strand break repair uncovered an unexpected nuclear function for this cytoskeletal protein.\",\n      \"evidence\": \"Quantitative mass spectrometry, siRNA knockdown, γ-irradiation, chromatin fractionation, immunofluorescence for repair foci, flow cytometry\",\n      \"pmids\": [\"36852271\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Whether MAP7D1 localizes to DNA damage sites itself or acts indirectly through cytoplasmic transport is unclear\",\n        \"Direct versus bridged interactions with RAD50 and 53BP1 have not been dissected\",\n        \"Relevance to other DNA repair pathways beyond NHEJ in G1 is untested\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"A patient-derived R201W mutation in the microtubule-binding domain demonstrated that loss of MAP7D1–microtubule interaction causes mitotic spindle instability, chromosome segregation errors, and RPS14 accumulation, linking MAP7D1 function to mitotic fidelity.\",\n      \"evidence\": \"Patient fibroblast analysis, microtubule co-sedimentation assay, mutant overexpression and siRNA depletion in T98G and HEK293T cells\",\n      \"pmids\": [\"40856631\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"The clinical syndrome associated with the R201W mutation has not been fully delineated\",\n        \"How MAP7D1 loss leads to RPS14 accumulation mechanistically is unexplained\",\n        \"Whether spindle defects arise from loss of kinesin-1 recruitment or a distinct mechanism is not resolved\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Showing that MAP7D1 selectively coats detyrosinated microtubules via expanded lattice recognition, recruits kinesin-1, and dynamically controls lysosome positioning in response to nutrient status provided a unified model for how the tubulin code directs organelle transport through MAP partitioning.\",\n      \"evidence\": \"(preprint) Live-cell imaging, rigor kinesin colocalization, MAP7D1 overexpression/knockdown, nutrient starvation/stimulation assays in cultured cells\",\n      \"pmids\": [\"bio_10.1101_2025.10.07.680844\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Findings are from a preprint not yet peer-reviewed\",\n        \"The structural basis for MAP7D1 preference for expanded detyrosinated lattice is not determined\",\n        \"Whether nutrient-dependent MAP7D1 density changes are regulated by signaling kinases or protein turnover is unknown\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"A unified structural and regulatory model explaining how MAP7D1 integrates its cytoskeletal, signaling, and nuclear functions — and whether these represent context-dependent moonlighting or a common mechanistic principle — remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No high-resolution structure of MAP7D1 bound to microtubules or interaction partners exists\",\n        \"How MAP7D1 shuttles between cytoplasmic and nuclear compartments is unknown\",\n        \"Genetic models in mammals with tissue-specific deletion are lacking\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [0, 1, 2, 5, 6]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [0, 1, 2, 5, 6]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [3]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [1]},\n      {\"term_id\": \"R-HSA-73894\", \"supporting_discovery_ids\": [3]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [5]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"DCLK1\",\n      \"DVL2\",\n      \"KIF5B\",\n      \"RAD50\",\n      \"53BP1\",\n      \"BRCA1\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```\n\n--- ROUND 2 (R2/R3, augmented corpus) ---\n\n```json\n{\n  \"mechanistic_narrative\": \"MAP7D1 is a microtubule-associated protein that functions as a microtubule-tethered activator of kinesin-1 and a regulator of microtubule stability, with roles spanning axon elongation, mitotic spindle integrity, DNA damage repair, and organelle positioning. MAP7D1 binds microtubules through its N-terminal domain and promotes kinesin-1 (KIF5B) landing and processivity both directly and allosterically through its C-terminal kinesin-binding domain, acting redundantly with MAP7 and MAP7D3 in HeLa cells [PMID:30770434]. DCLK1 phosphorylates MAP7D1 at Ser315 to drive callosal axon elongation in cortical neurons [PMID:27503845], while MAP7D1 also interacts with Disheveled to mediate Wnt5a-dependent cortical microtubule plus-end targeting via a kinesin-1-dependent feedback loop [PMID:29880710]. Beyond cytoskeletal functions, MAP7D1 participates in G1-phase DNA double-strand break repair by recruiting RAD50 and 53BP1 to damage sites [PMID:36852271], maintains acetylated stable microtubules in neuronal cells [PMID:35470240], and its loss of function causes multipolar spindles and lagging chromosomes during mitosis [PMID:40856631].\",\n  \"teleology\": [\n    {\n      \"year\": 2016,\n      \"claim\": \"Establishing MAP7D1 as a phosphorylation-dependent effector of axon elongation resolved how DCLK1 kinase activity is transduced into cytoskeletal remodeling during cortical neuron development.\",\n      \"evidence\": \"Proteomic substrate identification, in vitro kinase assay, and in utero electroporation with phosphomimetic rescue in cortical neurons\",\n      \"pmids\": [\"27503845\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Downstream mechanism by which Ser315 phosphorylation alters microtubule dynamics is unknown\",\n        \"Whether other kinases phosphorylate MAP7D1 at additional sites in neurons is untested\"\n      ]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Demonstrating that MAP7D1 bridges Disheveled and kinesin-1 on microtubules established a Wnt5a-responsive feedback loop for cortical microtubule plus-end targeting, revealing a signaling-to-transport coupling mechanism.\",\n      \"evidence\": \"Reciprocal co-immunoprecipitation, live-cell imaging in HeLa cells, depletion/rescue, and cross-species validation in Drosophila\",\n      \"pmids\": [\"29880710\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Structural basis of the MAP7D1–Disheveled interaction is unresolved\",\n        \"Physiological tissue context beyond cultured cells and Drosophila wings is unexplored\"\n      ]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Reconstitution of MAP7D1–kinesin-1 interactions with purified proteins demonstrated that MAP7 family members promote kinesin-1 microtubule landing through both direct tethering and allosteric activation, defining the core molecular mechanism of MAP7D1 as a motor cofactor.\",\n      \"evidence\": \"Single-molecule TIRF assays with purified proteins, domain truncation mapping, siRNA knockdown in HeLa cells\",\n      \"pmids\": [\"30770434\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Relative contributions of MAP7D1 versus MAP7 and MAP7D3 in specific tissues remain undefined\",\n        \"No high-resolution structure of the MAP7D1–kinesin-1 complex exists\"\n      ]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Zebrafish map7d1b knockout revealed that MAP7D1 supports autophagic flux and protein homeostasis in cardiac muscle, extending its functional repertoire beyond cytoskeletal regulation to organelle quality control.\",\n      \"evidence\": \"Zebrafish knockout, doxorubicin-induced cardiomyopathy model, autophagic flux and protein aggregation assays\",\n      \"pmids\": [\"34327238\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Whether autophagy impairment is a direct consequence of disrupted kinesin-1 transport or an independent mechanism is unclear\",\n        \"Mammalian cardiac phenotypes of MAP7D1 loss have not been reported\"\n      ]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Distinguishing MAP7D1's microtubule-stabilizing mechanism (via acetylation) from MAP7D2's direct binding-based stabilization clarified how paralog-specific functions diversify the microtubule cytoskeleton in neuronal cells.\",\n      \"evidence\": \"siRNA/shRNA knockdown, nocodazole resistance assay, acetylated/detyrosinated tubulin immunofluorescence in N1-E115 neuronal cells\",\n      \"pmids\": [\"35470240\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"How MAP7D1 promotes tubulin acetylation (e.g., via αTAT1 recruitment) is unknown\",\n        \"In vivo neuronal consequences of selective MAP7D1 loss on stable microtubule pools are untested\"\n      ]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Discovery that MAP7D1 recruits RAD50 and 53BP1 to DNA damage sites in G1 cells established an unexpected nuclear function for this microtubule-associated protein in the DNA double-strand break repair pathway.\",\n      \"evidence\": \"Quantitative AP-MS, γ-irradiation, chromatin fractionation, immunofluorescence, siRNA knockdown\",\n      \"pmids\": [\"36852271\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Whether MAP7D1 translocates to the nucleus or acts via an indirect cytoskeletal-signaling mechanism is unresolved\",\n        \"The DNA repair function has not been independently replicated\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Identification of a patient-derived MAP7D1 R201W mutation that disrupts microtubule binding and causes multipolar spindles and lagging chromosomes provided the first direct link between MAP7D1 dysfunction and mitotic fidelity defects in a disease context.\",\n      \"evidence\": \"Patient fibroblast analysis, mutant overexpression, siRNA knockdown, microtubule co-sedimentation, spindle morphology analysis in T98G and HEK293T cells\",\n      \"pmids\": [\"40856631\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Whether MAP7D1 R201W is causative for the patient's Shwachman-Diamond syndrome phenotype or a modifier is not established\",\n        \"Mechanism linking MAP7D1 loss to RPS14 accumulation in dividing cells is unexplained\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Showing that MAP7D1 preferentially partitions onto detyrosinated microtubules and that its density is nutrient-regulated to control lysosome positioning unified its kinesin-1 cofactor and tubulin-code reader activities into a nutrient-responsive transport mechanism.\",\n      \"evidence\": \"(preprint) Live-cell imaging, siRNA knockdown and overexpression, lysosome tracking, rigor kinesin localization assays\",\n      \"pmids\": [\"bio_10.1101_2025.10.07.680844\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Findings are from a preprint not yet peer-reviewed\",\n        \"Signaling pathway linking nutrient status to MAP7D1 density changes is unidentified\",\n        \"Whether detyrosination preference applies in vivo in specific tissues is untested\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"A high-resolution structure of MAP7D1 bound to microtubules and/or kinesin-1, the molecular basis of its nuclear DNA repair role, and how nutrient signals regulate MAP7D1–microtubule partitioning remain major unresolved questions.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"No structural data for MAP7D1 or its complexes exist\",\n        \"Nuclear versus cytoplasmic partitioning mechanism is unknown\",\n        \"Tissue-specific functional redundancy among MAP7 family paralogs is poorly defined\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [0, 1, 2, 3, 7, 8]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [0, 1, 2, 3, 7, 8]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [4]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [0, 1]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [7]},\n      {\"term_id\": \"R-HSA-73894\", \"supporting_discovery_ids\": [4]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [1]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"KIF5B\",\n      \"DVL2\",\n      \"DCLK1\",\n      \"RAD50\",\n      \"TP53BP1\",\n      \"BRCA1\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}