{"gene":"MYO1D","run_date":"2026-06-10T05:19:52","timeline":{"discoveries":[{"year":2000,"finding":"Myr4 (MYO1D) functions as an actin-based mechanoenzyme required for membrane trafficking along the recycling endosome pathway in MDCK epithelial cells. In vitro assays demonstrated that transfer of endocytosed markers from early endosomes to recycling endosomes depends on myr4 activity, its light chain calmodulin (CaM), and polymerized actin.","method":"In vitro endosomal transfer assay with functional perturbation of CaM and actin dynamics; identification of myr4 as CaM target protein","journal":"Traffic (Copenhagen, Denmark)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vitro functional assay with multiple perturbations (CaM inhibition, actin depolymerization), single lab, mechanistic conclusion supported by two orthogonal approaches","pmids":["11208135"],"is_preprint":false},{"year":2015,"finding":"Myo1D is required for planar cell polarity (PCP) in multi-ciliated tracheal and ependymal epithelial cells. In Myo1d knockout rats, rotational PCP of tracheal cilia is lost, cilia beat in a disordered pattern, and the asymmetric localization of PCP core protein Vangl1 is abolished. In ependymal cells, left-right positioning of basal body clusters is disrupted. Myo1d localizes to the F-actin and basal body-rich subapical cortex of ciliated tracheal cells.","method":"Knockout rat model (Myo1d KO), scanning and transmission electron microscopy, immunofluorescence localization, bead transport assays","journal":"Cytoskeleton (Hoboken, N.J.)","confidence":"High","confidence_rationale":"Tier 2 / Strong — clean KO with multiple defined phenotypic readouts (ciliary orientation, Vangl1 localization, bead transport) and direct localization experiment, single lab but multiple orthogonal methods","pmids":["26446290"],"is_preprint":false},{"year":2015,"finding":"Myo1D (Drosophila) functions with Nesprin1 (dNesp1) to direct mRNA transport from muscle nuclei to postsynaptic sites at the neuromuscular junction. Both dNesp1 and Myo1D are mutually required for their localization to immature boutons along F-actin-based 'railroad tracks.'","method":"Drosophila genetic analysis, fluorescence imaging of mRNA localization and protein co-localization, mutual requirement demonstrated by loss-of-function","journal":"Neuron","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic loss-of-function with defined phenotype (mRNA mislocalization, synaptic maturation defect) and reciprocal localization dependence; single lab","pmids":["25959729"],"is_preprint":false},{"year":2018,"finding":"Myosin1D (Myo1D) is an evolutionarily conserved regulator of left-right asymmetry. In zebrafish, Myo1D controls the formation and function of the left-right organizer (Kupffer's vesicle), regulates cilia orientation, and functionally interacts with the planar cell polarity pathway component VanGogh-like2 (Vangl2) to shape directional fluid flow for symmetry breaking.","method":"Zebrafish myo1d loss-of-function, genetic epistasis with vangl2, cilia orientation analysis, LRO flow measurements","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic loss-of-function with defined cellular and organ-level phenotypes, epistasis with Vangl2, multiple orthogonal readouts (cilia orientation, LRO flow, Nodal cascade), replication of concept across labs","pmids":["29769531"],"is_preprint":false},{"year":2018,"finding":"Myo1D is required for left-right axis formation in Xenopus. Morpholino-mediated myo1d knockdown disrupts organ laterality, impairs flow at the LR organizer through shorter, fewer, and non-polarized cilia, and inhibits convergent extension downstream of Wnt/PCP signaling. Genetic interaction between myo1d and vangl2 was demonstrated.","method":"Xenopus morpholino knockdown, organ placement scoring, Nodal cascade expression analysis, cilia imaging, gastrula explant convergent extension assay, ATF2 reporter for non-canonical Wnt, genetic epistasis with vangl2","journal":"Current biology : CB","confidence":"High","confidence_rationale":"Tier 2 / Strong — morpholino loss-of-function with multiple orthogonal readouts (organ placement, cilia phenotype, signaling reporters, convergent extension, genetic epistasis), independent replication of Myo1D-PCP functional interaction","pmids":["29478852"],"is_preprint":false},{"year":2019,"finding":"MYO1D anchors unphosphorylated EGFR family members (EGFR, ErbB2, ErbB4, but not ErbB3) to the plasma membrane by binding their kinase domains via a C-terminal β-meander motif in the MYO1D tail domain. MYO1D tethers these receptors to the underlying actin cytoskeleton before ligand activation. Overexpressed MYO1D increases EGFR levels, promotes cancer cell motility and viability, and drives colorectal tumor progression in a syngeneic mouse model.","method":"Co-immunoprecipitation, co-localization studies, domain-deletion/mutagenesis of MYO1D tail, functional phenotype assays (cell motility, viability), syngeneic mouse tumor model","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 1-2 / Moderate — direct binding demonstrated by Co-IP with domain mapping and mutagenesis identifying the β-meander motif, supported by functional loss-of-function and in vivo tumor model; single lab but multiple orthogonal methods","pmids":["31420606"],"is_preprint":false},{"year":2012,"finding":"Myosin-1d interacts with and is co-expressed with aspartoacylase in the nervous system. Myosin-1d localizes along axons and in the myelin compartment of the mouse sciatic nerve, and is enriched in Purkinje and granule cell layers of cerebellum, expanding to myelinated axonal tracts upon onset of myelination.","method":"Co-immunoprecipitation (interaction with aspartoacylase), immunofluorescence localization in tissue sections","journal":"Brain research","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single Co-IP identifying aspartoacylase as interactor, localization without direct functional consequence demonstrated","pmids":["22284616"],"is_preprint":false},{"year":2014,"finding":"Myo1d is expressed in myelinating oligodendrocytes in the CNS, localizing to abaxonal and adaxonal (outer and inner cytoplasm-containing loop) regions of myelin. Expression increases during myelinogenesis and continues into adulthood.","method":"Immunohistochemistry with isoform-specific antibody, Western blot fractionation of myelin","journal":"Journal of neuroscience research","confidence":"Low","confidence_rationale":"Tier 3 / Weak — localization by immunostaining without direct functional consequence in this paper; single lab, descriptive","pmids":["24903835"],"is_preprint":false},{"year":2016,"finding":"Myo1d knockdown in cultured oligodendrocytes causes retraction of processes, degeneration of myelin-like membrane, apoptosis, and impairs intracellular transport of myelin proteolipid protein (PLP). Myo1d localizes at the leading edge of myelin-like membrane, colocalizing with actin filaments, CNPase, and partially with PLP.","method":"siRNA knockdown, immunofluorescence colocalization, morphological analysis, PLP transport assay in cultured oligodendrocytes","journal":"ASN neuro","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — siRNA loss-of-function with defined morphological and transport phenotypes, supported by colocalization; single lab, two orthogonal approaches","pmids":["27655972"],"is_preprint":false},{"year":2017,"finding":"Myo1d knockdown in vivo during remyelination after cuprizone-induced demyelination decreases myelin basic protein and PLP levels, increases pro-inflammatory microglia and astrocytes, decreases anti-inflammatory microglia, and increases caspase-3-positive oligodendrocytes, indicating Myo1d is required for the remyelination process and oligodendrocyte survival.","method":"In vivo siRNA injection via stereotaxy in cuprizone demyelination mouse model, immunofluorescence, cell counting","journal":"Neurochemical research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo loss-of-function with multiple defined phenotypic readouts (myelin proteins, inflammatory markers, cell survival); single lab, single method (siRNA + histology)","pmids":["28986688"],"is_preprint":false},{"year":2022,"finding":"SPAG6 interacts with MYO1D (demonstrated by immunoprecipitation and mass spectrometry). Overexpression of SPAG6 promotes translocation of MYO1D from cytosol to the cell membrane, thereby upregulating EGFR family expression and activating PI3K/AKT and ERK signaling to promote AML cell proliferation and migration.","method":"Co-immunoprecipitation, mass spectrometry, subcellular fractionation, overexpression and knockdown functional assays, xenograft mouse model","journal":"Blood advances","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP/MS identifies SPAG6-MYO1D interaction, fractionation shows MYO1D membrane translocation, functional rescue/knockdown experiments; single lab","pmids":["35667090"],"is_preprint":false},{"year":2020,"finding":"In Drosophila, Myo1D is required for the membrane localization of the initiator caspase Dronc in mature enterocytes. In Myo1d mutant midguts, apoptosis-induced proliferation (AiP) signals including ROS generation, hemocyte recruitment, and JNK signaling are impaired, and intestinal stem cell mitotic activity is reduced.","method":"Drosophila myo1d mutant analysis, live imaging of Dronc membrane localization, ROS detection, JNK reporter assay, hemocyte recruitment scoring, ISC mitosis quantification","journal":"Cell reports","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic loss-of-function with multiple orthogonal mechanistic readouts (Dronc localization, ROS, JNK, hemocyte recruitment, ISC proliferation); replicated concept from prior Drosophila studies in a new tissue context","pmids":["33238125"],"is_preprint":false},{"year":2022,"finding":"In Drosophila, F-actin remodeling driven by LIMK1 acts downstream of the caspase Dronc and depends on Myo1D to mediate ROS production and JNK activation during apoptosis-induced proliferation (AiP). Synergistic effects of Dronc and LIMK1 co-expression on F-actin accumulation, ROS, and JNK activation are abolished in myo1d mutants, placing Myo1D as a required mediator in the Dronc-LIMK1-F-actin-ROS-JNK axis.","method":"Drosophila genetic epistasis (myo1d mutant background), LIMK1/cofilin overexpression and knockdown, ROS detection, JNK reporter assay, F-actin staining","journal":"PLoS genetics","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic epistasis with multiple defined molecular readouts places Myo1D in a specific pathway node; multiple orthogonal methods; extends prior mechanistic framework","pmids":["36469525"],"is_preprint":false},{"year":2023,"finding":"Drosophila Myo1D has distinct ATPase kinetics compared to Myo1C: a 12.5-fold higher actin-activated steady-state ATPase rate, 8-fold higher MgADP release rate, and rate-limiting step is MgADP release (vs. phosphate release for Myo1C). Myo1D propels actin filaments at higher speeds than Myo1C in in vitro gliding assays and robustly transports 50 nm unilamellar vesicles along actin filaments, while Myo1C does not support vesicle transport.","method":"Transient kinetic assays (ATPase mechanism determination), in vitro actin gliding assay, in vitro vesicle transport assay","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — reconstituted in vitro enzymatic assays with transient kinetics, gliding assay, and vesicle transport assay; multiple orthogonal Tier 1 methods in single rigorous study","pmids":["37380077"],"is_preprint":false},{"year":2024,"finding":"Drosophila Myo1D (but not Myo1C) exhibits a higher proportion of fast-diffusing molecules on the plasma membrane of macrophages, as measured by single-molecule imaging. This distinct membrane diffusion behavior depends on both head and tail domains of each myosin and does not exert mutual influence between paralogs. Myo1D's unique membrane dynamics may underlie its role in dextral cell chirality.","method":"Single-molecule imaging on Drosophila macrophage plasma membrane, diffusion analysis","journal":"Genes to cells : devoted to molecular & cellular mechanisms","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct single-molecule localization experiment with domain dissection; single lab, novel method but limited to diffusion characterization","pmids":["38454557"],"is_preprint":false},{"year":2025,"finding":"MYO1D preferentially binds PDGFRα/β heterodimers. Knockdown of MYO1D leads to retention of PDGFRα/β heterodimers at the plasma membrane, resulting in increased ERK1/2 phosphorylation and increased cell proliferation, indicating MYO1D normally promotes PDGFRα/β heterodimer internalization away from the plasma membrane to limit ERK signaling.","method":"PDGFR dimer-specific interactome (bimolecular fluorescence complementation + proteomics), siRNA knockdown of MYO1D, receptor internalization imaging, ERK1/2 phosphorylation assay, proliferation assay","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 / Strong — preferential binding identified by dimer-specific interactome, functional consequence of MYO1D knockdown on receptor localization, ERK signaling, and proliferation established with multiple orthogonal methods; peer-reviewed publication","pmids":["40404618"],"is_preprint":false},{"year":2025,"finding":"In Drosophila macrophages, Myo1D directs clockwise circumferential F-actin flow, while Myo1C directs counterclockwise flow; both depend on Myosin2 (Myo2). In a modified in vitro motility assay at near-physiological actin concentrations, Myo1D triggers self-organization of a clockwise-rotating chiral F-actin ring with parallel, annular barbed-end polarity, whereas Myo1C induces random F-actin flow. This provides a molecular basis for how Myo1D establishes dextral cell and organ chirality.","method":"Live imaging of F-actin flow in Drosophila macrophages, in vitro motility assay with near-physiological actin concentrations, genetic perturbation of Myo2","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 1-2 / Moderate — in vitro reconstitution of chiral F-actin ring plus live-cell imaging; preprint not yet peer-reviewed, single lab","pmids":["bio_10.1101_2025.05.06.648335"],"is_preprint":true},{"year":2009,"finding":"Xenopus laevis Myo1D (XlMyo1d) protein is present in eggs and increases at early neurula through tadpole stages. mRNA is expressed in neural tube, pre-somitic mesoderm, somites, and cranial neural crest cells during migration; during somitogenesis, transcript localizes to a stripe overlapping the nuclear region of somites. A novel isoform-specific tail homology embedded within the TH1 domain was identified by sequence analysis.","method":"Western blot (developmental expression), whole-mount in situ hybridization (mRNA localization), sequence analysis","journal":"Development, growth & differentiation","confidence":"Low","confidence_rationale":"Tier 3 / Weak — descriptive localization and expression data; no direct functional experiment; single lab","pmids":["19382939"],"is_preprint":false},{"year":2025,"finding":"In Drosophila, Myo1D ensures survival of apoptosis-resistant DARE cells by preventing lethal effector caspase activation, while Myo7A/Crinkled promotes effector caspase activation in the same cells. This positions Myo1D as a molecular switch controlling the threshold of caspase-driven apoptosis in a radiation-induced regenerative context.","method":"Drosophila genetic loss-of-function analysis in wing imaginal disc after ionizing radiation, caspase activity reporters, genetic interaction with Myo7A/Crinkled","journal":"Nature communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic loss-of-function with caspase activity readout and genetic comparison with Myo7A; single lab, mechanistic placement in caspase regulation pathway","pmids":["41345088"],"is_preprint":false},{"year":2022,"finding":"miR-217-5p targets MYO1D in rat podocytes, as validated by luciferase reporter assay, qRT-PCR, and Western blotting. Myo1d is predicted to be involved in actin filament organization in podocytes, and its downregulation by miR-217-5p is associated with podocyte morphological changes including shrunken cells with abnormal actin cytoskeletons.","method":"Luciferase reporter assay, qRT-PCR, Western blot (miRNA-target validation)","journal":"Non-coding RNA","confidence":"Low","confidence_rationale":"Tier 3 / Weak — miRNA targeting validated by standard assays but direct mechanistic role of MYO1D protein in podocytes not experimentally established beyond target validation; single lab","pmids":["35736640"],"is_preprint":false}],"current_model":"MYO1D is an unconventional class I myosin that uses calmodulin as a light chain and links the actin cytoskeleton to membranes; it functions as an actin-based motor with high ATPase activity and MgADP-release as rate-limiting step, capable of propelling actin in directional (leftward/clockwise) circles and transporting vesicles along actin filaments. At the plasma membrane, MYO1D anchors unphosphorylated EGFR family receptors (via its tail β-meander motif) and PDGFRα/β heterodimers (promoting their internalization to limit ERK signaling) to the actin cytoskeleton, and promotes membrane trafficking through recycling endosomes. MYO1D is an evolutionarily conserved regulator of left-right asymmetry, acting through the planar cell polarity (PCP)/Vangl2 pathway to orient cilia at the left-right organizer in vertebrates and to establish dextral cell chirality through circumferential F-actin flow organization in Drosophila. It additionally controls basal body/cilia planar polarity in tracheal and ependymal epithelia, mediates Dronc caspase membrane localization to drive apoptosis-induced proliferation, supports oligodendrocyte myelin membrane dynamics and PLP transport, and directs mRNA transport to postsynaptic sites in Drosophila muscle."},"narrative":{"mechanistic_narrative":"MYO1D is an unconventional class I myosin that couples the actin cytoskeleton to membranes and acts as a conserved organizer of cellular and organismal chirality. As a motor, it has high actin-activated ATPase activity with MgADP release as the rate-limiting step, propels actin filaments rapidly, and uniquely transports unilamellar vesicles along actin filaments, distinguishing it functionally from the paralog Myo1C [PMID:37380077]. In epithelial cells it drives membrane trafficking along the recycling endosome pathway in a calmodulin- and F-actin-dependent manner [PMID:11208135]. A central role of MYO1D is the establishment of left-right asymmetry: in zebrafish and Xenopus it controls left-right organizer formation, cilia number and orientation, and directional fluid flow, acting through genetic interaction with the planar cell polarity component Vangl2 [PMID:29769531, PMID:29478852], and in multiciliated tracheal and ependymal epithelia it is required for rotational PCP, ciliary orientation, and asymmetric Vangl localization [PMID:26446290]. In Drosophila this chirality function is mechanistically resolved as MYO1D directing clockwise circumferential F-actin flow, organizing self-rotating chiral actin rings to set dextral cell chirality [PMID:bio_10.1101_2025.05.06.648335]. At the plasma membrane MYO1D tethers unphosphorylated EGFR-family receptors (EGFR, ErbB2, ErbB4) to the actin cytoskeleton via a C-terminal β-meander motif in its tail, with overexpression elevating EGFR and promoting tumor cell motility and progression [PMID:31420606], whereas it promotes internalization of PDGFRα/β heterodimers away from the membrane to limit ERK signaling and proliferation [PMID:40404618]. In Drosophila MYO1D additionally controls membrane localization of the initiator caspase Dronc to drive apoptosis-induced proliferation through a Dronc–LIMK1–F-actin–ROS–JNK axis [PMID:33238125, PMID:36469525] and sets the threshold for caspase-driven apoptosis in regenerating tissue [PMID:41345088]. It also supports oligodendrocyte myelin membrane dynamics and PLP transport [PMID:27655972] and directs nuclear-to-postsynaptic mRNA transport with Nesprin1 at the Drosophila neuromuscular junction [PMID:25959729].","teleology":[{"year":2000,"claim":"Established MYO1D as an actin-based mechanoenzyme functioning in membrane trafficking, answering whether this myosin participates in intracellular transport.","evidence":"In vitro endosomal transfer assay in MDCK cells with calmodulin and actin perturbation","pmids":["11208135"],"confidence":"Medium","gaps":["Did not resolve the directionality or kinetics of the motor","No identification of cargo adaptors on recycling endosomes"]},{"year":2015,"claim":"Linked MYO1D to planar cell polarity in ciliated epithelia, showing it organizes ciliary orientation and asymmetric PCP protein localization.","evidence":"Myo1d knockout rat with EM, immunofluorescence, and bead transport assays","pmids":["26446290"],"confidence":"High","gaps":["Molecular mechanism connecting MYO1D motor activity to Vangl localization unresolved","Whether the same mechanism operates in left-right organizer cilia not addressed here"]},{"year":2015,"claim":"Showed MYO1D cooperates with Nesprin1 to transport mRNA along F-actin tracks to postsynaptic sites, extending its motor role to RNA localization.","evidence":"Drosophila genetic loss-of-function and co-localization imaging at the neuromuscular junction","pmids":["25959729"],"confidence":"Medium","gaps":["Direct cargo-binding interaction between MYO1D and mRNP not demonstrated","Generality to other transcripts or tissues unknown"]},{"year":2018,"claim":"Established MYO1D as an evolutionarily conserved left-right asymmetry regulator acting through the PCP/Vangl2 pathway, resolving how it shapes symmetry-breaking flow.","evidence":"Zebrafish and Xenopus loss-of-function with cilia orientation, LRO flow, convergent extension, and vangl2 epistasis","pmids":["29769531","29478852"],"confidence":"High","gaps":["Biochemical nature of the MYO1D-Vangl2 interaction not defined","How motor chirality translates to organ laterality not mechanistically resolved at this stage"]},{"year":2019,"claim":"Defined a membrane-anchoring function: MYO1D tethers unphosphorylated EGFR-family receptors to actin via a tail β-meander motif, with pro-tumorigenic consequences.","evidence":"Co-IP, domain mapping/mutagenesis, motility/viability assays, and syngeneic mouse tumor model","pmids":["31420606"],"confidence":"High","gaps":["How receptor tethering is regulated dynamically not established","Whether anchoring is restored upon ligand activation unaddressed"]},{"year":2020,"claim":"Identified MYO1D as required for membrane localization of the initiator caspase Dronc, linking it to apoptosis-induced proliferation signaling.","evidence":"Drosophila mutant analysis with Dronc localization imaging, ROS, JNK reporter, and ISC mitosis quantification","pmids":["33238125"],"confidence":"High","gaps":["Direct physical interaction between MYO1D and Dronc not shown","Whether mammalian caspases are similarly regulated unknown"]},{"year":2022,"claim":"Placed MYO1D as a required node in the Dronc-LIMK1-F-actin-ROS-JNK axis driving apoptosis-induced proliferation.","evidence":"Drosophila genetic epistasis with LIMK1/cofilin manipulation, ROS, JNK reporter, and F-actin staining","pmids":["36469525"],"confidence":"High","gaps":["Mechanistic step at which MYO1D acts within the F-actin remodeling cascade not pinpointed"]},{"year":2022,"claim":"Showed SPAG6 drives MYO1D translocation to the membrane to upregulate EGFR-family signaling in AML, connecting MYO1D localization to oncogenic PI3K/AKT and ERK activation.","evidence":"Co-IP/MS, subcellular fractionation, overexpression/knockdown, and xenograft model","pmids":["35667090"],"confidence":"Medium","gaps":["How SPAG6 triggers MYO1D translocation mechanistically unresolved","Direct vs. indirect SPAG6-MYO1D binding not fully dissected"]},{"year":2023,"claim":"Resolved MYO1D's motor mechanism, distinguishing it kinetically from Myo1C and establishing it as a vesicle-transporting motor.","evidence":"Transient kinetic ATPase assays, in vitro actin gliding, and vesicle transport assays with Drosophila Myo1D","pmids":["37380077"],"confidence":"High","gaps":["Whether vertebrate MYO1D shares identical kinetics not tested","In vivo vesicle cargoes not identified"]},{"year":2024,"claim":"Showed MYO1D's distinct plasma-membrane diffusion behavior depends on both head and tail domains, potentially underlying its chirality function.","evidence":"Single-molecule imaging on Drosophila macrophage membranes with domain dissection","pmids":["38454557"],"confidence":"Medium","gaps":["Causal link between diffusion behavior and chirality not demonstrated","Membrane lipid or protein determinants of diffusion unknown"]},{"year":2025,"claim":"Provided a molecular basis for chirality by showing MYO1D directs clockwise F-actin flow and self-organizes a chiral rotating actin ring, opposite to Myo1C.","evidence":"Live imaging of macrophage F-actin flow and in vitro motility reconstitution at near-physiological actin (preprint)","pmids":["bio_10.1101_2025.05.06.648335"],"confidence":"Medium","gaps":["Preprint not yet peer-reviewed","How chiral F-actin flow is read out to orient PCP/Vangl in vivo not established"]},{"year":2025,"claim":"Refined the receptor-trafficking model by showing MYO1D promotes internalization of PDGFRα/β heterodimers to limit ERK signaling, contrasting with its EGFR-anchoring role.","evidence":"Dimer-specific interactome, MYO1D knockdown, receptor internalization imaging, ERK phosphorylation and proliferation assays","pmids":["40404618"],"confidence":"High","gaps":["How MYO1D selectivity for heterodimers vs. homodimers is achieved unclear","Reconciliation of anchoring vs. internalization functions across receptor families not resolved"]},{"year":null,"claim":"How MYO1D's chiral actin-organizing motor activity is mechanistically coupled to the PCP/Vangl2 machinery and to its receptor-trafficking functions in a single unifying model remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural model linking motor chirality to Vangl2 polarization","Vertebrate cargo and adaptor inventory incomplete","Integration of EGFR-anchoring, PDGFR-internalization, and chirality roles unestablished"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0003774","term_label":"cytoskeletal motor activity","supporting_discovery_ids":[13,16,0]},{"term_id":"GO:0140657","term_label":"ATP-dependent activity","supporting_discovery_ids":[13]},{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[13,16,0]},{"term_id":"GO:0140313","term_label":"molecular sequestering activity","supporting_discovery_ids":[5]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[5,14,15,10]},{"term_id":"GO:0005768","term_label":"endosome","supporting_discovery_ids":[0]},{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[1,8,16]},{"term_id":"GO:0005815","term_label":"microtubule organizing center","supporting_discovery_ids":[1]}],"pathway":[{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[3,4,1]},{"term_id":"R-HSA-5653656","term_label":"Vesicle-mediated transport","supporting_discovery_ids":[0,13]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[5,15,10]},{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[11,12,18]}],"complexes":[],"partners":["VANGL2","EGFR","ERBB2","ERBB4","PDGFRA","PDGFRB","SPAG6","CALM1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"O94832","full_name":"Unconventional myosin-Id","aliases":[],"length_aa":1006,"mass_kda":116.2,"function":"Unconventional myosin that functions as actin-based motor protein with ATPase activity (By similarity). Plays a role in endosomal protein trafficking, and especially in the transfer of cargo proteins from early to recycling endosomes (By similarity). Required for normal planar cell polarity in ciliated tracheal cells, for normal rotational polarity of cilia, and for coordinated, unidirectional ciliary movement in the trachea. Required for normal, polarized cilia organization in brain ependymal epithelial cells (By similarity)","subcellular_location":"Cytoplasm; Perikaryon; Cell projection, dendrite; Early endosome; Cytoplasm, cell cortex","url":"https://www.uniprot.org/uniprotkb/O94832/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/MYO1D","classification":"Not Classified","n_dependent_lines":2,"n_total_lines":1208,"dependency_fraction":0.0016556291390728477},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"CALM1","stoichiometry":0.2},{"gene":"CALM2","stoichiometry":0.2},{"gene":"CALM3","stoichiometry":0.2},{"gene":"CAPZB","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/MYO1D","total_profiled":1310},"omim":[{"mim_id":"606539","title":"MYOSIN ID; MYO1D","url":"https://www.omim.org/entry/606539"},{"mim_id":"606538","title":"MYOSIN IC; MYO1C","url":"https://www.omim.org/entry/606538"},{"mim_id":"601480","title":"MYOSIN IF; MYO1F","url":"https://www.omim.org/entry/601480"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Cytosol","reliability":"Approved"}],"tissue_specificity":"Group enriched","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"blood 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\"Myr4 (MYO1D) functions as an actin-based mechanoenzyme required for membrane trafficking along the recycling endosome pathway in MDCK epithelial cells. In vitro assays demonstrated that transfer of endocytosed markers from early endosomes to recycling endosomes depends on myr4 activity, its light chain calmodulin (CaM), and polymerized actin.\",\n      \"method\": \"In vitro endosomal transfer assay with functional perturbation of CaM and actin dynamics; identification of myr4 as CaM target protein\",\n      \"journal\": \"Traffic (Copenhagen, Denmark)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vitro functional assay with multiple perturbations (CaM inhibition, actin depolymerization), single lab, mechanistic conclusion supported by two orthogonal approaches\",\n      \"pmids\": [\"11208135\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Myo1D is required for planar cell polarity (PCP) in multi-ciliated tracheal and ependymal epithelial cells. In Myo1d knockout rats, rotational PCP of tracheal cilia is lost, cilia beat in a disordered pattern, and the asymmetric localization of PCP core protein Vangl1 is abolished. In ependymal cells, left-right positioning of basal body clusters is disrupted. Myo1d localizes to the F-actin and basal body-rich subapical cortex of ciliated tracheal cells.\",\n      \"method\": \"Knockout rat model (Myo1d KO), scanning and transmission electron microscopy, immunofluorescence localization, bead transport assays\",\n      \"journal\": \"Cytoskeleton (Hoboken, N.J.)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — clean KO with multiple defined phenotypic readouts (ciliary orientation, Vangl1 localization, bead transport) and direct localization experiment, single lab but multiple orthogonal methods\",\n      \"pmids\": [\"26446290\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Myo1D (Drosophila) functions with Nesprin1 (dNesp1) to direct mRNA transport from muscle nuclei to postsynaptic sites at the neuromuscular junction. Both dNesp1 and Myo1D are mutually required for their localization to immature boutons along F-actin-based 'railroad tracks.'\",\n      \"method\": \"Drosophila genetic analysis, fluorescence imaging of mRNA localization and protein co-localization, mutual requirement demonstrated by loss-of-function\",\n      \"journal\": \"Neuron\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic loss-of-function with defined phenotype (mRNA mislocalization, synaptic maturation defect) and reciprocal localization dependence; single lab\",\n      \"pmids\": [\"25959729\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Myosin1D (Myo1D) is an evolutionarily conserved regulator of left-right asymmetry. In zebrafish, Myo1D controls the formation and function of the left-right organizer (Kupffer's vesicle), regulates cilia orientation, and functionally interacts with the planar cell polarity pathway component VanGogh-like2 (Vangl2) to shape directional fluid flow for symmetry breaking.\",\n      \"method\": \"Zebrafish myo1d loss-of-function, genetic epistasis with vangl2, cilia orientation analysis, LRO flow measurements\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic loss-of-function with defined cellular and organ-level phenotypes, epistasis with Vangl2, multiple orthogonal readouts (cilia orientation, LRO flow, Nodal cascade), replication of concept across labs\",\n      \"pmids\": [\"29769531\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Myo1D is required for left-right axis formation in Xenopus. Morpholino-mediated myo1d knockdown disrupts organ laterality, impairs flow at the LR organizer through shorter, fewer, and non-polarized cilia, and inhibits convergent extension downstream of Wnt/PCP signaling. Genetic interaction between myo1d and vangl2 was demonstrated.\",\n      \"method\": \"Xenopus morpholino knockdown, organ placement scoring, Nodal cascade expression analysis, cilia imaging, gastrula explant convergent extension assay, ATF2 reporter for non-canonical Wnt, genetic epistasis with vangl2\",\n      \"journal\": \"Current biology : CB\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — morpholino loss-of-function with multiple orthogonal readouts (organ placement, cilia phenotype, signaling reporters, convergent extension, genetic epistasis), independent replication of Myo1D-PCP functional interaction\",\n      \"pmids\": [\"29478852\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"MYO1D anchors unphosphorylated EGFR family members (EGFR, ErbB2, ErbB4, but not ErbB3) to the plasma membrane by binding their kinase domains via a C-terminal β-meander motif in the MYO1D tail domain. MYO1D tethers these receptors to the underlying actin cytoskeleton before ligand activation. Overexpressed MYO1D increases EGFR levels, promotes cancer cell motility and viability, and drives colorectal tumor progression in a syngeneic mouse model.\",\n      \"method\": \"Co-immunoprecipitation, co-localization studies, domain-deletion/mutagenesis of MYO1D tail, functional phenotype assays (cell motility, viability), syngeneic mouse tumor model\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — direct binding demonstrated by Co-IP with domain mapping and mutagenesis identifying the β-meander motif, supported by functional loss-of-function and in vivo tumor model; single lab but multiple orthogonal methods\",\n      \"pmids\": [\"31420606\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Myosin-1d interacts with and is co-expressed with aspartoacylase in the nervous system. Myosin-1d localizes along axons and in the myelin compartment of the mouse sciatic nerve, and is enriched in Purkinje and granule cell layers of cerebellum, expanding to myelinated axonal tracts upon onset of myelination.\",\n      \"method\": \"Co-immunoprecipitation (interaction with aspartoacylase), immunofluorescence localization in tissue sections\",\n      \"journal\": \"Brain research\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single Co-IP identifying aspartoacylase as interactor, localization without direct functional consequence demonstrated\",\n      \"pmids\": [\"22284616\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Myo1d is expressed in myelinating oligodendrocytes in the CNS, localizing to abaxonal and adaxonal (outer and inner cytoplasm-containing loop) regions of myelin. Expression increases during myelinogenesis and continues into adulthood.\",\n      \"method\": \"Immunohistochemistry with isoform-specific antibody, Western blot fractionation of myelin\",\n      \"journal\": \"Journal of neuroscience research\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — localization by immunostaining without direct functional consequence in this paper; single lab, descriptive\",\n      \"pmids\": [\"24903835\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Myo1d knockdown in cultured oligodendrocytes causes retraction of processes, degeneration of myelin-like membrane, apoptosis, and impairs intracellular transport of myelin proteolipid protein (PLP). Myo1d localizes at the leading edge of myelin-like membrane, colocalizing with actin filaments, CNPase, and partially with PLP.\",\n      \"method\": \"siRNA knockdown, immunofluorescence colocalization, morphological analysis, PLP transport assay in cultured oligodendrocytes\",\n      \"journal\": \"ASN neuro\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — siRNA loss-of-function with defined morphological and transport phenotypes, supported by colocalization; single lab, two orthogonal approaches\",\n      \"pmids\": [\"27655972\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Myo1d knockdown in vivo during remyelination after cuprizone-induced demyelination decreases myelin basic protein and PLP levels, increases pro-inflammatory microglia and astrocytes, decreases anti-inflammatory microglia, and increases caspase-3-positive oligodendrocytes, indicating Myo1d is required for the remyelination process and oligodendrocyte survival.\",\n      \"method\": \"In vivo siRNA injection via stereotaxy in cuprizone demyelination mouse model, immunofluorescence, cell counting\",\n      \"journal\": \"Neurochemical research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo loss-of-function with multiple defined phenotypic readouts (myelin proteins, inflammatory markers, cell survival); single lab, single method (siRNA + histology)\",\n      \"pmids\": [\"28986688\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"SPAG6 interacts with MYO1D (demonstrated by immunoprecipitation and mass spectrometry). Overexpression of SPAG6 promotes translocation of MYO1D from cytosol to the cell membrane, thereby upregulating EGFR family expression and activating PI3K/AKT and ERK signaling to promote AML cell proliferation and migration.\",\n      \"method\": \"Co-immunoprecipitation, mass spectrometry, subcellular fractionation, overexpression and knockdown functional assays, xenograft mouse model\",\n      \"journal\": \"Blood advances\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP/MS identifies SPAG6-MYO1D interaction, fractionation shows MYO1D membrane translocation, functional rescue/knockdown experiments; single lab\",\n      \"pmids\": [\"35667090\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"In Drosophila, Myo1D is required for the membrane localization of the initiator caspase Dronc in mature enterocytes. In Myo1d mutant midguts, apoptosis-induced proliferation (AiP) signals including ROS generation, hemocyte recruitment, and JNK signaling are impaired, and intestinal stem cell mitotic activity is reduced.\",\n      \"method\": \"Drosophila myo1d mutant analysis, live imaging of Dronc membrane localization, ROS detection, JNK reporter assay, hemocyte recruitment scoring, ISC mitosis quantification\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic loss-of-function with multiple orthogonal mechanistic readouts (Dronc localization, ROS, JNK, hemocyte recruitment, ISC proliferation); replicated concept from prior Drosophila studies in a new tissue context\",\n      \"pmids\": [\"33238125\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"In Drosophila, F-actin remodeling driven by LIMK1 acts downstream of the caspase Dronc and depends on Myo1D to mediate ROS production and JNK activation during apoptosis-induced proliferation (AiP). Synergistic effects of Dronc and LIMK1 co-expression on F-actin accumulation, ROS, and JNK activation are abolished in myo1d mutants, placing Myo1D as a required mediator in the Dronc-LIMK1-F-actin-ROS-JNK axis.\",\n      \"method\": \"Drosophila genetic epistasis (myo1d mutant background), LIMK1/cofilin overexpression and knockdown, ROS detection, JNK reporter assay, F-actin staining\",\n      \"journal\": \"PLoS genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic epistasis with multiple defined molecular readouts places Myo1D in a specific pathway node; multiple orthogonal methods; extends prior mechanistic framework\",\n      \"pmids\": [\"36469525\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Drosophila Myo1D has distinct ATPase kinetics compared to Myo1C: a 12.5-fold higher actin-activated steady-state ATPase rate, 8-fold higher MgADP release rate, and rate-limiting step is MgADP release (vs. phosphate release for Myo1C). Myo1D propels actin filaments at higher speeds than Myo1C in in vitro gliding assays and robustly transports 50 nm unilamellar vesicles along actin filaments, while Myo1C does not support vesicle transport.\",\n      \"method\": \"Transient kinetic assays (ATPase mechanism determination), in vitro actin gliding assay, in vitro vesicle transport assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — reconstituted in vitro enzymatic assays with transient kinetics, gliding assay, and vesicle transport assay; multiple orthogonal Tier 1 methods in single rigorous study\",\n      \"pmids\": [\"37380077\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Drosophila Myo1D (but not Myo1C) exhibits a higher proportion of fast-diffusing molecules on the plasma membrane of macrophages, as measured by single-molecule imaging. This distinct membrane diffusion behavior depends on both head and tail domains of each myosin and does not exert mutual influence between paralogs. Myo1D's unique membrane dynamics may underlie its role in dextral cell chirality.\",\n      \"method\": \"Single-molecule imaging on Drosophila macrophage plasma membrane, diffusion analysis\",\n      \"journal\": \"Genes to cells : devoted to molecular & cellular mechanisms\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct single-molecule localization experiment with domain dissection; single lab, novel method but limited to diffusion characterization\",\n      \"pmids\": [\"38454557\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"MYO1D preferentially binds PDGFRα/β heterodimers. Knockdown of MYO1D leads to retention of PDGFRα/β heterodimers at the plasma membrane, resulting in increased ERK1/2 phosphorylation and increased cell proliferation, indicating MYO1D normally promotes PDGFRα/β heterodimer internalization away from the plasma membrane to limit ERK signaling.\",\n      \"method\": \"PDGFR dimer-specific interactome (bimolecular fluorescence complementation + proteomics), siRNA knockdown of MYO1D, receptor internalization imaging, ERK1/2 phosphorylation assay, proliferation assay\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — preferential binding identified by dimer-specific interactome, functional consequence of MYO1D knockdown on receptor localization, ERK signaling, and proliferation established with multiple orthogonal methods; peer-reviewed publication\",\n      \"pmids\": [\"40404618\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"In Drosophila macrophages, Myo1D directs clockwise circumferential F-actin flow, while Myo1C directs counterclockwise flow; both depend on Myosin2 (Myo2). In a modified in vitro motility assay at near-physiological actin concentrations, Myo1D triggers self-organization of a clockwise-rotating chiral F-actin ring with parallel, annular barbed-end polarity, whereas Myo1C induces random F-actin flow. This provides a molecular basis for how Myo1D establishes dextral cell and organ chirality.\",\n      \"method\": \"Live imaging of F-actin flow in Drosophila macrophages, in vitro motility assay with near-physiological actin concentrations, genetic perturbation of Myo2\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — in vitro reconstitution of chiral F-actin ring plus live-cell imaging; preprint not yet peer-reviewed, single lab\",\n      \"pmids\": [\"bio_10.1101_2025.05.06.648335\"],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Xenopus laevis Myo1D (XlMyo1d) protein is present in eggs and increases at early neurula through tadpole stages. mRNA is expressed in neural tube, pre-somitic mesoderm, somites, and cranial neural crest cells during migration; during somitogenesis, transcript localizes to a stripe overlapping the nuclear region of somites. A novel isoform-specific tail homology embedded within the TH1 domain was identified by sequence analysis.\",\n      \"method\": \"Western blot (developmental expression), whole-mount in situ hybridization (mRNA localization), sequence analysis\",\n      \"journal\": \"Development, growth & differentiation\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — descriptive localization and expression data; no direct functional experiment; single lab\",\n      \"pmids\": [\"19382939\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"In Drosophila, Myo1D ensures survival of apoptosis-resistant DARE cells by preventing lethal effector caspase activation, while Myo7A/Crinkled promotes effector caspase activation in the same cells. This positions Myo1D as a molecular switch controlling the threshold of caspase-driven apoptosis in a radiation-induced regenerative context.\",\n      \"method\": \"Drosophila genetic loss-of-function analysis in wing imaginal disc after ionizing radiation, caspase activity reporters, genetic interaction with Myo7A/Crinkled\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic loss-of-function with caspase activity readout and genetic comparison with Myo7A; single lab, mechanistic placement in caspase regulation pathway\",\n      \"pmids\": [\"41345088\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"miR-217-5p targets MYO1D in rat podocytes, as validated by luciferase reporter assay, qRT-PCR, and Western blotting. Myo1d is predicted to be involved in actin filament organization in podocytes, and its downregulation by miR-217-5p is associated with podocyte morphological changes including shrunken cells with abnormal actin cytoskeletons.\",\n      \"method\": \"Luciferase reporter assay, qRT-PCR, Western blot (miRNA-target validation)\",\n      \"journal\": \"Non-coding RNA\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — miRNA targeting validated by standard assays but direct mechanistic role of MYO1D protein in podocytes not experimentally established beyond target validation; single lab\",\n      \"pmids\": [\"35736640\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"MYO1D is an unconventional class I myosin that uses calmodulin as a light chain and links the actin cytoskeleton to membranes; it functions as an actin-based motor with high ATPase activity and MgADP-release as rate-limiting step, capable of propelling actin in directional (leftward/clockwise) circles and transporting vesicles along actin filaments. At the plasma membrane, MYO1D anchors unphosphorylated EGFR family receptors (via its tail β-meander motif) and PDGFRα/β heterodimers (promoting their internalization to limit ERK signaling) to the actin cytoskeleton, and promotes membrane trafficking through recycling endosomes. MYO1D is an evolutionarily conserved regulator of left-right asymmetry, acting through the planar cell polarity (PCP)/Vangl2 pathway to orient cilia at the left-right organizer in vertebrates and to establish dextral cell chirality through circumferential F-actin flow organization in Drosophila. It additionally controls basal body/cilia planar polarity in tracheal and ependymal epithelia, mediates Dronc caspase membrane localization to drive apoptosis-induced proliferation, supports oligodendrocyte myelin membrane dynamics and PLP transport, and directs mRNA transport to postsynaptic sites in Drosophila muscle.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"MYO1D is an unconventional class I myosin that couples the actin cytoskeleton to membranes and acts as a conserved organizer of cellular and organismal chirality. As a motor, it has high actin-activated ATPase activity with MgADP release as the rate-limiting step, propels actin filaments rapidly, and uniquely transports unilamellar vesicles along actin filaments, distinguishing it functionally from the paralog Myo1C [#13]. In epithelial cells it drives membrane trafficking along the recycling endosome pathway in a calmodulin- and F-actin-dependent manner [#0]. A central role of MYO1D is the establishment of left-right asymmetry: in zebrafish and Xenopus it controls left-right organizer formation, cilia number and orientation, and directional fluid flow, acting through genetic interaction with the planar cell polarity component Vangl2 [#3, #4], and in multiciliated tracheal and ependymal epithelia it is required for rotational PCP, ciliary orientation, and asymmetric Vangl localization [#1]. In Drosophila this chirality function is mechanistically resolved as MYO1D directing clockwise circumferential F-actin flow, organizing self-rotating chiral actin rings to set dextral cell chirality [#16]. At the plasma membrane MYO1D tethers unphosphorylated EGFR-family receptors (EGFR, ErbB2, ErbB4) to the actin cytoskeleton via a C-terminal β-meander motif in its tail, with overexpression elevating EGFR and promoting tumor cell motility and progression [#5], whereas it promotes internalization of PDGFRα/β heterodimers away from the membrane to limit ERK signaling and proliferation [#15]. In Drosophila MYO1D additionally controls membrane localization of the initiator caspase Dronc to drive apoptosis-induced proliferation through a Dronc–LIMK1–F-actin–ROS–JNK axis [#11, #12] and sets the threshold for caspase-driven apoptosis in regenerating tissue [#18]. It also supports oligodendrocyte myelin membrane dynamics and PLP transport [#8] and directs nuclear-to-postsynaptic mRNA transport with Nesprin1 at the Drosophila neuromuscular junction [#2].\",\n  \"teleology\": [\n    {\n      \"year\": 2000,\n      \"claim\": \"Established MYO1D as an actin-based mechanoenzyme functioning in membrane trafficking, answering whether this myosin participates in intracellular transport.\",\n      \"evidence\": \"In vitro endosomal transfer assay in MDCK cells with calmodulin and actin perturbation\",\n      \"pmids\": [\"11208135\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Did not resolve the directionality or kinetics of the motor\", \"No identification of cargo adaptors on recycling endosomes\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Linked MYO1D to planar cell polarity in ciliated epithelia, showing it organizes ciliary orientation and asymmetric PCP protein localization.\",\n      \"evidence\": \"Myo1d knockout rat with EM, immunofluorescence, and bead transport assays\",\n      \"pmids\": [\"26446290\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Molecular mechanism connecting MYO1D motor activity to Vangl localization unresolved\", \"Whether the same mechanism operates in left-right organizer cilia not addressed here\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Showed MYO1D cooperates with Nesprin1 to transport mRNA along F-actin tracks to postsynaptic sites, extending its motor role to RNA localization.\",\n      \"evidence\": \"Drosophila genetic loss-of-function and co-localization imaging at the neuromuscular junction\",\n      \"pmids\": [\"25959729\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Direct cargo-binding interaction between MYO1D and mRNP not demonstrated\", \"Generality to other transcripts or tissues unknown\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Established MYO1D as an evolutionarily conserved left-right asymmetry regulator acting through the PCP/Vangl2 pathway, resolving how it shapes symmetry-breaking flow.\",\n      \"evidence\": \"Zebrafish and Xenopus loss-of-function with cilia orientation, LRO flow, convergent extension, and vangl2 epistasis\",\n      \"pmids\": [\"29769531\", \"29478852\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Biochemical nature of the MYO1D-Vangl2 interaction not defined\", \"How motor chirality translates to organ laterality not mechanistically resolved at this stage\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Defined a membrane-anchoring function: MYO1D tethers unphosphorylated EGFR-family receptors to actin via a tail β-meander motif, with pro-tumorigenic consequences.\",\n      \"evidence\": \"Co-IP, domain mapping/mutagenesis, motility/viability assays, and syngeneic mouse tumor model\",\n      \"pmids\": [\"31420606\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"How receptor tethering is regulated dynamically not established\", \"Whether anchoring is restored upon ligand activation unaddressed\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Identified MYO1D as required for membrane localization of the initiator caspase Dronc, linking it to apoptosis-induced proliferation signaling.\",\n      \"evidence\": \"Drosophila mutant analysis with Dronc localization imaging, ROS, JNK reporter, and ISC mitosis quantification\",\n      \"pmids\": [\"33238125\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Direct physical interaction between MYO1D and Dronc not shown\", \"Whether mammalian caspases are similarly regulated unknown\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Placed MYO1D as a required node in the Dronc-LIMK1-F-actin-ROS-JNK axis driving apoptosis-induced proliferation.\",\n      \"evidence\": \"Drosophila genetic epistasis with LIMK1/cofilin manipulation, ROS, JNK reporter, and F-actin staining\",\n      \"pmids\": [\"36469525\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Mechanistic step at which MYO1D acts within the F-actin remodeling cascade not pinpointed\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Showed SPAG6 drives MYO1D translocation to the membrane to upregulate EGFR-family signaling in AML, connecting MYO1D localization to oncogenic PI3K/AKT and ERK activation.\",\n      \"evidence\": \"Co-IP/MS, subcellular fractionation, overexpression/knockdown, and xenograft model\",\n      \"pmids\": [\"35667090\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"How SPAG6 triggers MYO1D translocation mechanistically unresolved\", \"Direct vs. indirect SPAG6-MYO1D binding not fully dissected\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Resolved MYO1D's motor mechanism, distinguishing it kinetically from Myo1C and establishing it as a vesicle-transporting motor.\",\n      \"evidence\": \"Transient kinetic ATPase assays, in vitro actin gliding, and vesicle transport assays with Drosophila Myo1D\",\n      \"pmids\": [\"37380077\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Whether vertebrate MYO1D shares identical kinetics not tested\", \"In vivo vesicle cargoes not identified\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Showed MYO1D's distinct plasma-membrane diffusion behavior depends on both head and tail domains, potentially underlying its chirality function.\",\n      \"evidence\": \"Single-molecule imaging on Drosophila macrophage membranes with domain dissection\",\n      \"pmids\": [\"38454557\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Causal link between diffusion behavior and chirality not demonstrated\", \"Membrane lipid or protein determinants of diffusion unknown\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Provided a molecular basis for chirality by showing MYO1D directs clockwise F-actin flow and self-organizes a chiral rotating actin ring, opposite to Myo1C.\",\n      \"evidence\": \"Live imaging of macrophage F-actin flow and in vitro motility reconstitution at near-physiological actin (preprint)\",\n      \"pmids\": [\"bio_10.1101_2025.05.06.648335\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Preprint not yet peer-reviewed\", \"How chiral F-actin flow is read out to orient PCP/Vangl in vivo not established\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Refined the receptor-trafficking model by showing MYO1D promotes internalization of PDGFRα/β heterodimers to limit ERK signaling, contrasting with its EGFR-anchoring role.\",\n      \"evidence\": \"Dimer-specific interactome, MYO1D knockdown, receptor internalization imaging, ERK phosphorylation and proliferation assays\",\n      \"pmids\": [\"40404618\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"How MYO1D selectivity for heterodimers vs. homodimers is achieved unclear\", \"Reconciliation of anchoring vs. internalization functions across receptor families not resolved\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How MYO1D's chiral actin-organizing motor activity is mechanistically coupled to the PCP/Vangl2 machinery and to its receptor-trafficking functions in a single unifying model remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"No structural model linking motor chirality to Vangl2 polarization\", \"Vertebrate cargo and adaptor inventory incomplete\", \"Integration of EGFR-anchoring, PDGFR-internalization, and chirality roles unestablished\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0003774\", \"supporting_discovery_ids\": [13, 16, 0]},\n      {\"term_id\": \"GO:0140657\", \"supporting_discovery_ids\": [13]},\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [13, 16, 0]},\n      {\"term_id\": \"GO:0140313\", \"supporting_discovery_ids\": [5]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [5, 14, 15, 10]},\n      {\"term_id\": \"GO:0005768\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [1, 8, 16]},\n      {\"term_id\": \"GO:0005815\", \"supporting_discovery_ids\": [1]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [3, 4, 1]},\n      {\"term_id\": \"R-HSA-5653656\", \"supporting_discovery_ids\": [0, 13]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [5, 15, 10]},\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [11, 12, 18]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"VANGL2\", \"EGFR\", \"ERBB2\", \"ERBB4\", \"PDGFRA\", \"PDGFRB\", \"SPAG6\", \"CALM1\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}