title: Nuclear localization of protein kinase C-alpha induces thyroid hormone receptor-alpha1 expression in the cardiomyocyte.
abstract: Maladaptive cardiac hypertrophy results in phenotypic changes in several genes that are thyroid hormone responsive, suggesting that thyroid hormone receptor (TR) function may be altered by cellular kinases, including protein kinase C (PKC) isozymes that are activated in pathological hypertrophy. To investigate the role of PKC signaling in regulating TR function, cultured neonatal rat ventricular myocytes were transduced with adenovirus (Ad) expressing wild-type (wt) or kinase-inactive (dn) PKC alpha or constitutively active (ca) PKC delta and PKC epsilon. Overexpression of wtPKC alpha, but not caPKC delta or caPKC epsilon, induced a 28-fold increase (P < 0.001) in TR alpha1 protein in the nuclear compartment and a smaller increase in the cytosol. Furthermore, TR alpha1 mRNA was increased 55-fold (P < 0.001). This effect of PKC alpha was dependent on its kinase activity because dnPKC alpha was without effect. Phorbol 12-myristate 13-acetate (PMA) induced nuclear translocation of endogenous PKC alpha and Ad-wtPKC alpha concomitantly with an increase in nuclear TR alpha1 protein. In contrast, PMA-induced nuclear translocation of dnPKC alpha resulted in a decrease of TR alpha1. The increase in TR alpha1 protein in Ad-wtPKC alpha-transduced cardiomyocytes was not the result of a reduced rate of protein degradation, nor was the half-life of TR alpha1 mRNA prolonged, suggesting a PKC alpha-mediated effect on TR alpha transcription. Although phosphorylation of ERK1/2 was increased in Ad-wtPKC alpha-transduced cells, inhibition of phospho-ERK did not change TR alpha1 expression. PKC alpha overexpression in cardiomyocytes caused marked repression of triiodothyronine (T3)-responsive genes, alpha-myosin heavy chain, and the sarcoplasmic reticulum calcium-activated adenosinetriphosphatase SERCA2. Treatment with T3 for 4 h resulted in significant reductions of PKC alpha in nuclear and cytosolic compartments, and decreased TR alpha1 mRNA and protein, with normalization of phenotype. These results implicate PKC alpha as a regulator of TR function and suggest that nuclear localization of PKC alpha may control transcription of the TR alpha gene, and consequently, affect cardiac phenotype.
authors.affiliation: Institute for Medical Research, North Shore-LIJ Health System, 350 Community Dr., Manhasset, NY 11030, USA.
authors.forename: Agnes
authors.initials: A
authors.lastname: Kenessey
authors.forename: Elizabeth Ann
authors.initials: EA
authors.lastname: Sullivan
authors.forename: Kaie
authors.initials: K
authors.lastname: Ojamaa
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D000831:Animals, Newborn; D000252:Calcium-Transporting ATPases / Q000096:biosynthesis; D002467:Cell Nucleus / Q000201:enzymology; D005786:Gene Expression Regulation; D019950:Mitogen-Activated Protein Kinase 1 / Q000502:physiology; D048052:Mitogen-Activated Protein Kinase 3 / Q000502:physiology; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000378:metabolism*; D018995:Myosin Heavy Chains / Q000096:biosynthesis; D019913:Nuclear Localization Signals; D010641:Phenotype; D051571:Protein Kinase C-alpha / Q000378:metabolism*; D051381:Rats; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D037021:Thyroid Hormone Receptors alpha / Q000096:biosynthesis*; D014161:Transduction, Genetic; D014284:Triiodothyronine / Q000494:pharmacology
chemical_list: C506948:Atp2a2 protein, rat; D019913:Nuclear Localization Signals; D037021:Thyroid Hormone Receptors alpha; D014284:Triiodothyronine; D051571:Protein Kinase C-alpha; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases; D018995:Myosin Heavy Chains; D000252:Calcium-Transporting ATPases; D013755:Tetradecanoylphorbol Acetate