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Constitutive phosphorylation of cardiac myosin regulatory light chain prevents development of hypertrophic cardiomyopathy in mice

Journal Article · · Proceedings of the National Academy of Sciences of the United States of America
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  1. Univ. of Miami Miller School of Medicine, FL (United States). Dept. of Molecular and Cellular Pharmacology
  2. Illinois Inst. of Technology, Chicago, IL (United States). Dept. of Biological, Chemical and Physical Sciences
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Myosin light chain kinase (MLCK)-dependent phosphorylation of the regulatory light chain (RLC) of cardiac myosin is known to play a beneficial role in heart disease, but the idea of a phosphorylation-mediated reversal of a hypertrophic cardiomyopathy (HCM) phenotype is novel. Our previous studies on transgenic (Tg) HCM-RLC mice revealed that the D166V (Aspartate166 →Valine) mutation-induced changes in heart morphology and function coincided with largely reduced RLC phosphorylation in situ. In this paper, we hypothesized that the introduction of a constitutively phosphorylated Serine15 (S15D) into the hearts of D166V mice would prevent the development of a deleterious HCM phenotype. In support of this notion, MLCK-induced phosphorylation of D166V-mutated hearts was found to rescue some of their abnormal contractile properties. Tg-S15D-D166V mice were generated with the human cardiac RLC-S15D-D166V construct substituted for mouse cardiac RLC and were subjected to functional, structural, and morphological assessments. The results were compared with Tg-WT and Tg-D166V mice expressing the human ventricular RLC-WT or its D166V mutant, respectively. Echocardiography and invasive hemodynamic studies demonstrated significant improvements of intact heart function in S15D-D166V mice compared with D166V, with the systolic and diastolic indices reaching those monitored in WT mice. A largely reduced maximal tension and abnormally high myofilament Ca2+ sensitivity observed in D166V-mutated hearts were reversed in S15D-D166V mice. Low-angle X-ray diffraction study revealed that altered myofilament structures present in HCM-D166V mice were mitigated in S15D-D166V rescue mice. Finally, our collective results suggest that expression of pseudophosphorylated RLC in the hearts of HCM mice is sufficient to prevent the development of the pathological HCM phenotype.
Research Organization:
Univ. of Miami Miller School of Medicine, FL (United States)
Sponsoring Organization:
American Heart Association (United States); National Inst. of Health (NIH) (United States); USDOE Office of Science (SC)
Contributing Organization:
Illinois Inst. of Technology, Chicago, IL (United States)
Grant/Contract Number:
AC02-06CH11357
OSTI ID:
1212938
Journal Information:
Proceedings of the National Academy of Sciences of the United States of America, Journal Name: Proceedings of the National Academy of Sciences of the United States of America Journal Issue: 30 Vol. 112; ISSN 0027-8424
Publisher:
National Academy of Sciences, Washington, DC (United States)Copyright Statement
Country of Publication:
United States
Language:
ENGLISH

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Cited By (13)

Phosphorylation of the regulatory light chain of myosin in striated muscle: methodological perspectives journal April 2016
Regulatory light chain phosphorylation augments length-dependent contraction in PTU-treated rats journal December 2018
A Novel Method of Determining the Functional Effects of a Minor Genetic Modification of a Protein journal November 2015
Molecular and Functional Effects of a Splice Site Mutation in the MYL2 Gene Associated with Cardioskeletal Myopathy and Early Cardiac Death in Infants journal June 2016
Direct Sarcomere Modulators Are Promising New Treatments for Cardiomyopathies journal December 2019
Effects of myosin variants on interacting-heads motif explain distinct hypertrophic and dilated cardiomyopathy phenotypes journal June 2017
Therapeutic potential of AAV9-S15D-RLC gene delivery in humanized MYL2 mouse model of HCM journal May 2019
Hereditary heart disease: pathophysiology, clinical presentation, and animal models of HCM, RCM, and DCM associated with mutations in cardiac myosin light chains journal January 2019
Insights into myosin regulatory and essential light chains: a focus on their roles in cardiac and skeletal muscle function, development and disease journal May 2019
Cardiac myosin light chain is phosphorylated by Ca 2+ /calmodulin-dependent and -independent kinase activities journal June 2016
Sarcomeric perturbations of myosin motors lead to dilated cardiomyopathy in genetically modified MYL2 mice journal February 2018
Hypercontractile mutant of ventricular myosin essential light chain leads to disruption of sarcomeric structure and function and results in restrictive cardiomyopathy in mice journal March 2017
Revisiting Frank-Starling: regulatory light chain phosphorylation alters the rate of force redevelopment ( k tr ) in a length-dependent fashion: RLC phosphorylation alters k tr in rat trabeculae journal July 2016

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Related Subjects

60 APPLIED LIFE SCIENCES
X-ray structure
cardiomyopathy
hemodynamics
myocardial contraction
myosin RLC