Lu, FujianMa, QingXie, WenjunLiou, Carter L.Zhang, DonghuiSweat, Mason E.Jardin, Blake D.Naya, Francisco J.Guo, YuxuanCheng, HepingPu, William T.2023-07-252023-07-252022-04-21F. Lu, Q. Ma, W. Xie, C.L. Liou, D. Zhang, M.E. Sweat, B.D. Jardin, F.J. Naya, Y. Guo, H. Cheng, W.T. Pu. 2022. "CMYA5 establishes cardiac dyad architecture and positioning." Nature Communications, Volume 13, Issue 1, pp.2185-. https://doi.org/10.1038/s41467-022-29902-42041-1723https://hdl.handle.net/2144/46493Cardiac excitation-contraction coupling requires dyads, the nanoscopic microdomains formed adjacent to Z-lines by apposition of transverse tubules and junctional sarcoplasmic reticulum. Disruption of dyad architecture and function are common features of diseased cardiomyocytes. However, little is known about the mechanisms that modulate dyad organization during cardiac development, homeostasis, and disease. Here, we use proximity proteomics in intact, living hearts to identify proteins enriched near dyads. Among these proteins is CMYA5, an under-studied striated muscle protein that co-localizes with Z-lines, junctional sarcoplasmic reticulum proteins, and transverse tubules in mature cardiomyocytes. During cardiac development, CMYA5 positioning adjacent to Z-lines precedes junctional sarcoplasmic reticulum positioning or transverse tubule formation. CMYA5 ablation disrupts dyad architecture, dyad positioning at Z-lines, and junctional sarcoplasmic reticulum Ca2+ release, leading to cardiac dysfunction and inability to tolerate pressure overload. These data provide mechanistic insights into cardiomyopathy pathogenesis by demonstrating that CMYA5 anchors junctional sarcoplasmic reticulum to Z-lines, establishes dyad architecture, and regulates dyad Ca2+ release.p. 2185ElectronicenAttribution 4.0 InternationalThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutoryregulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.http://creativecommons.org/licenses/by/4.0/http://creativecommons.org/licenses/by/4.0/CalciumExcitation contraction couplingMuscle proteinsMyocytes, cardiacSarcolemmaSarcoplasmic reticulumArticle2023-02-0910.1038/s41467-022-29902-40000-0002-1376-5235 (Jardin, Blake D)0000-0002-9604-6702 (Cheng, Heping)745290