{"gene":"MYLK3","run_date":"2026-04-29T11:37:56","timeline":{"discoveries":[{"year":2020,"finding":"A null mutation in Mylk3 in C57BL/6N mice abolishes MYLK3 protein expression and causes dilated cardiomyopathy with eccentric hypertrophy and defects in sarcomere organisation, establishing MYLK3 as essential for cardiac function.","method":"RNAseq variant calling, immunofluorescent staining of cardiomyocytes, mouse genetic model (Mylk3 null)","journal":"Life science alliance","confidence":"High","confidence_rationale":"Tier 2 — clean KO with defined cardiac phenotype, multiple orthogonal methods (RNAseq, immunofluorescence, echocardiography)","pmids":["32213617"],"is_preprint":false},{"year":2017,"finding":"MYLK3 loss-of-function mutations (read-through c.2459A>C and frameshift c.1879_1885del) in humans result in markedly reduced MYLK3 protein expression and reduced myosin light chain 2 (MLC2) phosphorylation, causing familial dilated cardiomyopathy.","method":"Whole exome sequencing with segregation analysis, in vitro expression experiments, immunohistochemistry for MLC2 phosphorylation","journal":"Scientific reports","confidence":"High","confidence_rationale":"Tier 2 — human genetics combined with in vitro functional validation of phosphorylation loss, consistent with mouse and zebrafish loss-of-function models","pmids":["29235529"],"is_preprint":false},{"year":2019,"finding":"miR-200c directly targets the MYLK3/MLCK gene in cardiomyocytes; increased miR-200c reduces MLCK and its downstream effector p-MLC2, while miR-200c inhibition increases MLCK and p-MLC2 expression, mediating cardiac hypertrophy.","method":"miRNA mimic/inhibitor transfection in neonatal rat cardiomyocytes, Western blotting, qRT-PCR, aortic banding rat model","journal":"Journal of cellular and molecular medicine","confidence":"Medium","confidence_rationale":"Tier 2 — loss/gain-of-function miRNA with defined downstream phosphorylation readout, single lab","pmids":["30680929"],"is_preprint":false}],"current_model":"MYLK3 (cardiac myosin light chain kinase) phosphorylates myosin regulatory light chain 2 (MLC2) in cardiomyocytes; loss of MYLK3 protein expression—whether through spontaneous null mutation in mice or loss-of-function mutations in humans—abolishes MLC2 phosphorylation and causes dilated cardiomyopathy with sarcomere disorganisation, and its expression is post-transcriptionally regulated by miR-200c."},"narrative":{"teleology":[{"year":2017,"claim":"Establishing MYLK3 as a human disease gene: it was unknown whether MYLK3 loss of function causes cardiomyopathy in humans; whole-exome sequencing of familial dilated cardiomyopathy kindreds identified two distinct loss-of-function MYLK3 mutations that abolished protein expression and MLC2 phosphorylation, directly linking MYLK3 kinase activity to human cardiac disease.","evidence":"Whole exome sequencing with segregation analysis, in vitro expression of mutant constructs, immunohistochemistry for phospho-MLC2 in patient myocardium","pmids":["29235529"],"confidence":"High","gaps":["Structural basis for how the read-through and frameshift mutations disrupt kinase function is not defined","Whether residual kinase-dead MYLK3 protein exerts dominant-negative effects versus simple haploinsufficiency is unresolved","No therapeutic rescue experiments were performed"]},{"year":2019,"claim":"Defining post-transcriptional regulation of MYLK3: it was unclear how MYLK3 expression is modulated in cardiac stress; miR-200c was shown to directly target MYLK3 mRNA, and its manipulation bidirectionally controlled MYLK3 and p-MLC2 levels, establishing a regulatory axis linking a stress-responsive miRNA to sarcomere phosphorylation.","evidence":"miRNA mimic/inhibitor transfection in neonatal rat cardiomyocytes with Western blotting and qRT-PCR readouts; aortic banding rat hypertrophy model","pmids":["30680929"],"confidence":"Medium","gaps":["Direct binding of miR-200c to the MYLK3 3′-UTR was not validated by luciferase reporter assay in this study","Whether miR-200c regulation of MYLK3 operates in adult human cardiomyocytes is untested","Contribution of miR-200c–MYLK3 axis relative to other hypertrophy pathways is not quantified"]},{"year":2020,"claim":"Confirming MYLK3 essentiality in vivo: the question of whether complete MYLK3 loss is sufficient to cause cardiac failure was resolved by characterizing a spontaneous Mylk3-null mouse, which developed dilated cardiomyopathy with eccentric hypertrophy and sarcomere disorganization, validating the human genetic findings in a mammalian model.","evidence":"RNAseq variant calling, immunofluorescence of cardiomyocyte sarcomeres, echocardiography in C57BL/6N Mylk3-null mice","pmids":["32213617"],"confidence":"High","gaps":["Whether cardiac-restricted re-expression of MYLK3 rescues the phenotype has not been tested","Specific MLC2 phosphorylation sites affected by MYLK3 loss were not mapped in this study","Temporal progression from MYLK3 loss to sarcomere disorganization is not characterized"]},{"year":null,"claim":"Key unresolved questions include the structural basis for MYLK3 substrate selectivity, whether MYLK3 phosphorylates additional cardiac substrates beyond MLC2, and whether pharmacological augmentation of MYLK3 activity or expression can reverse established dilated cardiomyopathy.","evidence":"","pmids":[],"confidence":"Low","gaps":["No crystal structure or cryo-EM model of MYLK3 exists","Full substrate repertoire of MYLK3 in the heart has not been determined by unbiased phosphoproteomics","No gain-of-function rescue study has been reported in any cardiomyopathy model"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[0,1]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[0,1]}],"pathway":[{"term_id":"R-HSA-397014","term_label":"Muscle contraction","supporting_discovery_ids":[0,1]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[1]}],"complexes":[],"partners":["MYL2"],"other_free_text":[]},"mechanistic_narrative":"MYLK3 is a cardiac-specific myosin light chain kinase that phosphorylates myosin regulatory light chain 2 (MLC2) and is essential for sarcomere organization and normal cardiac function [PMID:29235529, PMID:32213617]. Loss-of-function mutations in MYLK3 in humans cause familial dilated cardiomyopathy with markedly reduced MLC2 phosphorylation [PMID:29235529], and a null mutation in mice recapitulates this phenotype with eccentric hypertrophy and sarcomere disorganization [PMID:32213617]. MYLK3 expression in cardiomyocytes is post-transcriptionally repressed by miR-200c, which reduces p-MLC2 levels and contributes to cardiac hypertrophy [PMID:30680929]."},"prefetch_data":{"uniprot":{"accession":"Q32MK0","full_name":"Myosin light chain kinase 3","aliases":["Cardiac-MyBP-C-associated Ca/CaM kinase","Cardiac-MLCK"],"length_aa":819,"mass_kda":88.4,"function":"Kinase that phosphorylates MYL2 in vitro. Promotes sarcomere formation in cardiomyocytes and increases cardiomyocyte contractility (By similarity)","subcellular_location":"Cytoplasm","url":"https://www.uniprot.org/uniprotkb/Q32MK0/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/MYLK3","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/MYLK3","total_profiled":1310},"omim":[{"mim_id":"612147","title":"MYOSIN LIGHT CHAIN KINASE 3; MYLK3","url":"https://www.omim.org/entry/612147"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Nucleoplasm","reliability":"Approved"},{"location":"Vesicles","reliability":"Additional"},{"location":"Cytosol","reliability":"Additional"}],"tissue_specificity":"Group enriched","tissue_distribution":"Detected in some","driving_tissues":[{"tissue":"heart muscle","ntpm":96.8},{"tissue":"skeletal muscle","ntpm":43.4},{"tissue":"tongue","ntpm":67.7}],"url":"https://www.proteinatlas.org/search/MYLK3"},"hgnc":{"alias_symbol":["caMLCK","MLCK"],"prev_symbol":[]},"alphafold":{"accession":"Q32MK0","domains":[{"cath_id":"3.30.200.20","chopping":"508-592","consensus_level":"medium","plddt":93.188,"start":508,"end":592},{"cath_id":"1.10.510.10","chopping":"593-793","consensus_level":"medium","plddt":91.976,"start":593,"end":793}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q32MK0","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q32MK0-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q32MK0-F1-predicted_aligned_error_v6.png","plddt_mean":61.72},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=MYLK3","jax_strain_url":"https://www.jax.org/strain/search?query=MYLK3"},"sequence":{"accession":"Q32MK0","fasta_url":"https://rest.uniprot.org/uniprotkb/Q32MK0.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q32MK0/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q32MK0"}},"corpus_meta":[{"pmid":"14743221","id":"PMC_14743221","title":"FAK-Src signalling through paxillin, ERK and MLCK regulates adhesion disassembly.","date":"2004","source":"Nature cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/14743221","citation_count":1144,"is_preprint":false},{"pmid":"23619146","id":"PMC_23619146","title":"TNFR2 activates MLCK-dependent tight junction dysregulation to cause apoptosis-mediated barrier loss and experimental colitis.","date":"2013","source":"Gastroenterology","url":"https://pubmed.ncbi.nlm.nih.gov/23619146","citation_count":311,"is_preprint":false},{"pmid":"17476691","id":"PMC_17476691","title":"Roles of Rho/ROCK and MLCK in TNF-alpha-induced changes in endothelial morphology and permeability.","date":"2007","source":"Journal of cellular physiology","url":"https://pubmed.ncbi.nlm.nih.gov/17476691","citation_count":219,"is_preprint":false},{"pmid":"20404178","id":"PMC_20404178","title":"MLCK-dependent exchange and actin binding region-dependent anchoring of ZO-1 regulate tight junction barrier function.","date":"2010","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/20404178","citation_count":207,"is_preprint":false},{"pmid":"32863203","id":"PMC_32863203","title":"Targeting NOX4 alleviates sepsis-induced acute lung injury via attenuation of redox-sensitive activation of CaMKII/ERK1/2/MLCK and endothelial cell barrier dysfunction.","date":"2020","source":"Redox biology","url":"https://pubmed.ncbi.nlm.nih.gov/32863203","citation_count":201,"is_preprint":false},{"pmid":"11350763","id":"PMC_11350763","title":"Stress fiber organization regulated by MLCK and Rho-kinase in cultured human fibroblasts.","date":"2001","source":"American journal of physiology. 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\"High\",\n      \"confidence_rationale\": \"Tier 2 — clean KO with defined cardiac phenotype, multiple orthogonal methods (RNAseq, immunofluorescence, echocardiography)\",\n      \"pmids\": [\"32213617\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"MYLK3 loss-of-function mutations (read-through c.2459A>C and frameshift c.1879_1885del) in humans result in markedly reduced MYLK3 protein expression and reduced myosin light chain 2 (MLC2) phosphorylation, causing familial dilated cardiomyopathy.\",\n      \"method\": \"Whole exome sequencing with segregation analysis, in vitro expression experiments, immunohistochemistry for MLC2 phosphorylation\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — human genetics combined with in vitro functional validation of phosphorylation loss, consistent with mouse and zebrafish loss-of-function models\",\n      \"pmids\": [\"29235529\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"miR-200c directly targets the MYLK3/MLCK gene in cardiomyocytes; increased miR-200c reduces MLCK and its downstream effector p-MLC2, while miR-200c inhibition increases MLCK and p-MLC2 expression, mediating cardiac hypertrophy.\",\n      \"method\": \"miRNA mimic/inhibitor transfection in neonatal rat cardiomyocytes, Western blotting, qRT-PCR, aortic banding rat model\",\n      \"journal\": \"Journal of cellular and molecular medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — loss/gain-of-function miRNA with defined downstream phosphorylation readout, single lab\",\n      \"pmids\": [\"30680929\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"MYLK3 (cardiac myosin light chain kinase) phosphorylates myosin regulatory light chain 2 (MLC2) in cardiomyocytes; loss of MYLK3 protein expression—whether through spontaneous null mutation in mice or loss-of-function mutations in humans—abolishes MLC2 phosphorylation and causes dilated cardiomyopathy with sarcomere disorganisation, and its expression is post-transcriptionally regulated by miR-200c.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"MYLK3 is a cardiac-specific myosin light chain kinase that phosphorylates myosin regulatory light chain 2 (MLC2) and is essential for sarcomere organization and normal cardiac function [PMID:29235529, PMID:32213617]. Loss-of-function mutations in MYLK3 in humans cause familial dilated cardiomyopathy with markedly reduced MLC2 phosphorylation [PMID:29235529], and a null mutation in mice recapitulates this phenotype with eccentric hypertrophy and sarcomere disorganization [PMID:32213617]. MYLK3 expression in cardiomyocytes is post-transcriptionally repressed by miR-200c, which reduces p-MLC2 levels and contributes to cardiac hypertrophy [PMID:30680929].\",\n  \"teleology\": [\n    {\n      \"year\": 2017,\n      \"claim\": \"Establishing MYLK3 as a human disease gene: it was unknown whether MYLK3 loss of function causes cardiomyopathy in humans; whole-exome sequencing of familial dilated cardiomyopathy kindreds identified two distinct loss-of-function MYLK3 mutations that abolished protein expression and MLC2 phosphorylation, directly linking MYLK3 kinase activity to human cardiac disease.\",\n      \"evidence\": \"Whole exome sequencing with segregation analysis, in vitro expression of mutant constructs, immunohistochemistry for phospho-MLC2 in patient myocardium\",\n      \"pmids\": [\"29235529\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Structural basis for how the read-through and frameshift mutations disrupt kinase function is not defined\",\n        \"Whether residual kinase-dead MYLK3 protein exerts dominant-negative effects versus simple haploinsufficiency is unresolved\",\n        \"No therapeutic rescue experiments were performed\"\n      ]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Defining post-transcriptional regulation of MYLK3: it was unclear how MYLK3 expression is modulated in cardiac stress; miR-200c was shown to directly target MYLK3 mRNA, and its manipulation bidirectionally controlled MYLK3 and p-MLC2 levels, establishing a regulatory axis linking a stress-responsive miRNA to sarcomere phosphorylation.\",\n      \"evidence\": \"miRNA mimic/inhibitor transfection in neonatal rat cardiomyocytes with Western blotting and qRT-PCR readouts; aortic banding rat hypertrophy model\",\n      \"pmids\": [\"30680929\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Direct binding of miR-200c to the MYLK3 3′-UTR was not validated by luciferase reporter assay in this study\",\n        \"Whether miR-200c regulation of MYLK3 operates in adult human cardiomyocytes is untested\",\n        \"Contribution of miR-200c–MYLK3 axis relative to other hypertrophy pathways is not quantified\"\n      ]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Confirming MYLK3 essentiality in vivo: the question of whether complete MYLK3 loss is sufficient to cause cardiac failure was resolved by characterizing a spontaneous Mylk3-null mouse, which developed dilated cardiomyopathy with eccentric hypertrophy and sarcomere disorganization, validating the human genetic findings in a mammalian model.\",\n      \"evidence\": \"RNAseq variant calling, immunofluorescence of cardiomyocyte sarcomeres, echocardiography in C57BL/6N Mylk3-null mice\",\n      \"pmids\": [\"32213617\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Whether cardiac-restricted re-expression of MYLK3 rescues the phenotype has not been tested\",\n        \"Specific MLC2 phosphorylation sites affected by MYLK3 loss were not mapped in this study\",\n        \"Temporal progression from MYLK3 loss to sarcomere disorganization is not characterized\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Key unresolved questions include the structural basis for MYLK3 substrate selectivity, whether MYLK3 phosphorylates additional cardiac substrates beyond MLC2, and whether pharmacological augmentation of MYLK3 activity or expression can reverse established dilated cardiomyopathy.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No crystal structure or cryo-EM model of MYLK3 exists\",\n        \"Full substrate repertoire of MYLK3 in the heart has not been determined by unbiased phosphoproteomics\",\n        \"No gain-of-function rescue study has been reported in any cardiomyopathy model\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [0, 1]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [0, 1]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-397014\", \"supporting_discovery_ids\": [0, 1]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [1]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"MYL2\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}