title: Cyclic mechanical strain of myocytes modifies CapZβ1 post translationally via PKCε.
abstract: The heart is exquisitely sensitive to mechanical stimuli and adapts to increased demands for work by enlarging the cardiomyocytes. In order to determine links between mechano-transduction mechanisms and hypertrophy, neonatal rat ventricular myocytes (NRVM) were subjected to physiologic strain for analysis of the dynamics of the actin capping protein, CapZ, and its post-translational modifications (PTM). CapZ binding rates were assessed after strain by fluorescence recovery after photobleaching (FRAP) of green fluorescent protein (GFP) expressed by a GFP-CapZβ1 adenovirus. To assess the role of the protein kinase C epsilon isoform (PKCε), rest or cyclic strain were combined with specific PKCε activation by constitutively active PKCε, or by inhibition with dominant negative PKCε (dnPKCε) expression. Significant increases of CapZ FRAP kinetics with strain were blunted by dnPKCε, suggesting that PKCε is involved in mechano-transduction signaling. Similar combinations of strain and PKC regulation in NRVMs were studied by PTM profiles of CapZβ1 using quantitative two-dimensional gel electrophoresis. The significantly increased charge on CapZ seen with mechanical strain was reversed by the addition of dnPKCε. Potential clinical relevance was confirmed in vivo by PTMs of CapZ in the failing heart of one-year old transgenic mice over-expressing PKCε. Furthermore, with strain there was significant PKCε translocation to the Z-disc and co-localization with CapZβ1 or α-actinin, which was quantified on confocal images. A hypothetical model is presented proposing that one destination of the mechanotransduction signaling pathways might be for PTMs of CapZ thereby regulating actin capping and filament assembly.
authors.affiliation: Department of Physiology and Biophysics, College of Medicine, University of Illinois at Chicago, MC 901, 835 S. Wolcott, Chicago, IL, 60612, USA.
authors.forename: Ying-Hsi
authors.initials: YH
authors.lastname: Lin
authors.affiliation: Department of Physiology and Biophysics, College of Medicine, University of Illinois at Chicago, MC 901, 835 S. Wolcott, Chicago, IL, 60612, USA.
authors.forename: Erik R
authors.initials: ER
authors.lastname: Swanson
authors.affiliation: Department of Physiology and Biophysics, College of Medicine, University of Illinois at Chicago, MC 901, 835 S. Wolcott, Chicago, IL, 60612, USA.
authors.forename: Jieli
authors.initials: J
authors.lastname: Li
authors.affiliation: Department of Physiology and Biophysics, College of Medicine, University of Illinois at Chicago, MC 901, 835 S. Wolcott, Chicago, IL, 60612, USA.|Department of Bioengineering, College of Engineering, University of Illinois at Chicago, Chicago, IL, 60607, USA.
authors.forename: Michael A
authors.initials: MA
authors.lastname: Mkrtschjan
authors.affiliation: Department of Physiology and Biophysics, College of Medicine, University of Illinois at Chicago, MC 901, 835 S. Wolcott, Chicago, IL, 60612, USA. russell@uic.edu.|Department of Bioengineering, College of Engineering, University of Illinois at Chicago, Chicago, IL, 60607, USA. russell@uic.edu.
authors.forename: Brenda
authors.identifier: http://orcid.org/0000-0002-4847-5836
authors.initials: B
authors.lastname: Russell
mesh_terms: D000185:Actinin / Q000235:genetics / Q000378:metabolism; D000818:Animals; D051347:CapZ Actin Capping Protein / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism*; D051744:Protein Kinase C-epsilon / Q000235:genetics / Q000378:metabolism*; D011499:Protein Processing, Post-Translational / Q000502:physiology*; D021381:Protein Transport / Q000502:physiology; D051381:Rats; D013314:Stress, Mechanical
chemical_list: D051347:CapZ Actin Capping Protein; D000185:Actinin; C497256:Prkce protein, mouse; C497257:Prkce protein, rat; D051744:Protein Kinase C-epsilon
keywords: Actin capping; Mechano-signaling; Mechano-transduction; Proteomics; Sarcomere