S165F mutation of junctophilin 2 affects Ca2+ signalling in skeletal muscle.

Abstract

JPs (junctophilins) contribute to the formation of junctional membrane complexes in muscle cells by physically linking the t-tubule (transverse-tubule) and SR (sarcoplasmic reticulum) membranes. In humans with HCM (hypertrophic cardiomyopathy), mutations in JP2 are linked to altered Ca²⁺ signalling in cardiomyocytes; however, the effects of these mutations on skeletal muscle function have not been examined. In the present study, we investigated the role of the dominant-negative JP2-S165F mutation (which is associated with human HCM) in skeletal muscle.

Consistent with the hypertrophy observed in human cardiac muscle, overexpression of JP2-S165F in primary mouse skeletal myotubes led to a significant increase in myotube diameter and resting cytosolic Ca²⁺ concentration. Single myotube Ca²⁺ imaging experiments showed reductions in both the excitation–contraction coupling gain and RyR (ryanodine receptor) 1-mediated Ca²⁺ release from the SR.

Immunoprecipitation assays revealed defects in the PKC (protein kinase C)-mediated phosphorylation of the JP2-S165F mutant protein at Ser165 and in binding of JP2-S165F to the Ca²⁺ channel TRPC3 (transient receptor potential cation canonical-type channel 3) on the t-tubule membrane. Therefore both the hypertrophy and altered intracellular Ca²⁺ signalling in the JP2-S165F-expressing skeletal myotubes can be linked to altered phosphorylation of JP2 and/or altered cross-talk among Ca²⁺ channels on the t-tubule and SR membranes.

Citation

Woo, J.S., Hwang, J.H., Ko, J.K., Weisleder, N., Ma, J. and Lee, E.H., 2010. S165F mutation of junctophilin 2 affects Ca2+ signalling in skeletal muscle. Biochemical Journal, 427(1), pp.125-134.

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Category: Medical & Pharmaceutical

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