Cardiomyocyte external mechanical unloading activates modifications of alpha-actinin differently from sarcomere-originated unloading

ABSTRACT: Loss of myocardial mass in a neonatal rat cardiomyocyte culture is studied to determine whether there is a distinguishable cellular response based on the origin of mechano-signals. The approach herein compares the sarcomeric assembly and disassembly processes in heart cells by imposing mechano-signals at the interface with the extracellular matrix (extrinsic) and at the level of the myofilaments (intrinsic). Experiments compared the effects of imposed internal (inside/out) and external (outside/in) loading and unloading on modifications in neonatal rat cardiomyocytes. Unloading of the cellular substrate by myosin inhibition (1uM mavacamten), or cessation of cyclic strain (1 Hz, 10% strain) after preconditioning, led to significant disassembly of sarcomeric alpha-actinin by 6 hrs. In myosin inhibition, this was accompanied by redistribution of intracellular poly-ubiquitin K48 to the cellular periphery relative to the poly-ubiquitin K48 reservoir at the I-band. Moreover, loading and unloading of the cellular substrate led to a three-fold increase in post translational modifications (PTMs) when compared to the myosin-specific activation or inhibition. Specifically, phosphorylation increased with loading while ubiquitination increased with unloading, which may involve ERK1/2 and FAK activation. The identified PTMs, including ubiquitination, acetylation, and phosphorylation, are proposed to modify internal domains in alpha-actinin to increase its propensity to bind F-actin. These results demonstrate a link between mechanical feedback and sarcomere protein homeostasis via PTMs of alpha-actinin that exemplify how cardiomyocytes exhibit differential responses to the origin of force. The implications of sarcomere regulation governed by PTMs of alpha-actinin are discussed with respect to cardiac atrophy and heart failure.

INSTRUMENT(S): timsTOF Pro

ORGANISM(S): Rattus Norvegicus (ncbitaxon:10116)

SUBMITTER: Brenda Russell  

PROVIDER: MSV000091010 | MassIVE | Thu Jan 05 06:39:00 GMT 2023

SECONDARY ACCESSION(S): PXD039284

REPOSITORIES: MassIVE