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Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2024,11,12]]},"abstract":"<jats:p>\n                    Skeletal muscle actin (ACTA1) mutations are a prevalent cause of skeletal myopathies consistent with ACTA1\u2019s high expression in skeletal muscle. Rare de novo mutations in ACTA1 associated with combined cardiac and skeletal myopathies have been reported, but ACTA1 represents only ~20% of the total actin pool in cardiomyocytes, making its role in cardiomyopathy controversial. Here we demonstrate how a mutation in an actin isoform expressed at low levels in cardiomyocytes can cause cardiomyopathy by focusing on a unique ACTA1 variant, R256H. We previously identified this variant in a family with dilated cardiomyopathy, who had reduced systolic function without clinical skeletal myopathy. Using a battery of multiscale biophysical tools, we show that R256H has potent effects on ACTA1 function at the molecular scale and in human cardiomyocytes. Importantly, we demonstrate that R256H acts in a dominant manner, where the incorporation of small amounts of mutant protein into thin filaments is sufficient to disrupt molecular contractility, and that this effect is dependent on the presence of troponin and tropomyosin. To understand the structural basis of this change in regulation, we resolved a structure of R256H filaments using cryoelectron microscopy, and we see alterations in actin\u2019s structure that have the potential to disrupt interactions with tropomyosin. Finally, we show that\n                    <jats:italic>\n                      ACTA1\n                      <jats:sup>R256H\/+<\/jats:sup>\n                    <\/jats:italic>\n                    human-induced pluripotent stem cell cardiomyocytes demonstrate reduced contractility and sarcomeric organization. Taken together, we demonstrate that R256H has multiple effects on ACTA1 function that are sufficient to cause reduced contractility and establish a likely causative relationship between ACTA1 R256H and clinical cardiomyopathy.\n                  <\/jats:p>","DOI":"10.1073\/pnas.2405020121","type":"journal-article","created":{"date-parts":[[2024,11,6]],"date-time":"2024-11-06T11:02:37Z","timestamp":1730890957000},"update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":13,"title":["Dilated cardiomyopathy\u2013associated skeletal muscle actin (ACTA1) mutation R256H disrupts actin structure and function and causes cardiomyocyte hypocontractility"],"prefix":"10.1073","volume":"121","author":[{"given":"Ankit","family":"Garg","sequence":"first","affiliation":[{"name":"Division of Cardiology, Department of Medicine, Johns Hopkins University, Baltimore, MD 21205"},{"name":"Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, MO 63110"},{"name":"Cardiovascular Division, Department of Medicine, Washington University School of Medicine, St. Louis, MO 63110"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5119-367X","authenticated-orcid":false,"given":"Silvia","family":"Jansen","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Physiology, Washington University in St. Louis, St. Louis, MO 63110"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lina","family":"Greenberg","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, MO 63110"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rui","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, MO 63110"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kory J.","family":"Lavine","sequence":"additional","affiliation":[{"name":"Cardiovascular Division, Department of Medicine, Washington University School of Medicine, St. Louis, MO 63110"},{"name":"Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1320-3547","authenticated-orcid":false,"given":"Michael J.","family":"Greenberg","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, MO 63110"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2024,11,6]]},"reference":[{"key":"e_1_3_4_1_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jchf.2020.03.014"},{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.116.309396"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jacc.2021.08.039"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jacbts.2023.07.013"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jacc.2016.08.079"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1260419"},{"key":"e_1_3_4_7_2","doi-asserted-by":"publisher","DOI":"10.3389\/fphys.2018.00405"},{"key":"e_1_3_4_8_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.nmd.2005.08.004"},{"key":"e_1_3_4_9_2","doi-asserted-by":"publisher","DOI":"10.1542\/peds.2012-1139"},{"key":"e_1_3_4_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jns.2018.08.015"},{"key":"e_1_3_4_11_2","doi-asserted-by":"publisher","DOI":"10.1002\/mgg3.2008"},{"key":"e_1_3_4_12_2","doi-asserted-by":"publisher","DOI":"10.1212\/WNL.0000000000010947"},{"key":"e_1_3_4_13_2","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCGEN.118.002243"},{"key":"e_1_3_4_14_2","first-page":"850","article-title":"A new mutation in the ACTA1 gene possibly associated with dilated cardiomyopathy without concomitant myopathy","volume":"75","author":"Diaz Exposito A.","year":"2022","unstructured":"A. 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Pardon Insights into single hiPSC-derived cardiomyocyte phenotypes and maturation using ConTraX an efficient pipeline for tracking contractile dynamics. bioRxiv [Preprint] (2021). https:\/\/doi.org\/10.1101\/2021.03.18.436014 (Accessed 3 January 2024).","DOI":"10.1101\/2021.03.18.436014"},{"key":"e_1_3_4_79_2","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.121.318868"},{"key":"e_1_3_4_80_2","doi-asserted-by":"crossref","unstructured":"A. Garg Data from \u201cDilated cardiomyopathy-associated skeletal muscle actin (ACTA1) mutation R256H disrupts actin structure and function and causes cardiomyocyte hypocontractility.\u201d Johns Hopkins Libraries. https:\/\/doi.org\/10.7281\/T1\/MX05CQ. Deposited 15 October 2024.","DOI":"10.1101\/2024.03.10.583979"},{"key":"e_1_3_4_81_2","doi-asserted-by":"crossref","unstructured":"A. Garg M. J. Greenberg R. Zhang Cryo-EM structure of recombinant wildtype ACTA1 phalloidin-stabilized F-actin. 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