Tetracycline-inducible CaM kinase II silences hypertrophy-sensitive gene expression in rat neonate cardiomyocytes.

Abstract

Recent work from this laboratory both in rat primary cardiomyocytes and in ventricular tissue of transgenic mouse models of induced hypertrophy has identified two Ca(2+)/calmodulin-dependent nuclear signaling cascades. The first involves the phosphatase calcineurin (CaN). The second is the CaM kinase kinase cascade which involves CaM kinase I and CaM kinase IV. Each of these signaling cascades strongly up-regulate transcription of hypertrophy-sensitive genes in the rat ventricular cardiomyocyte. We have documented that over-expression of an active form of CaM kinase II silenced transcriptional induction of hypertrophy-sensitive genes. The purpose of this study was to generate an inducible CaM kinase II expression system and correlate its expression with the silencing of hypertrophic-sensitive reporters. A truncated form of CaM KII, CaM KII (1-290) was subcloned downstream and proximal to a promoter under transcriptional control (induction) of the tetracycline-regulated transcription factor, tet-TransActivator (tTA). Hypertrophy-sensitive reporter activity in primary cardiomyocytes was silenced when tet-inducible CaM KII was co-expressed with plasmids harboring active forms of CaN, CaM KI or CaM KIV. For instance, induced CaM KII expression silenced CaN, CaM kinase I, or CaM kinase IV driven ANF reporter activity 4.9-, 2.9-, and 6.9-fold below their maximal values, respectively. Myocyte exposure to doxycycline (DOX) blocked tTA-driven CaM KII expression and restored CaN/CaM KI or CaN/CaM KIV driven reporter activation. This study demonstrates, for the first time, that active CaM KII silences Ca(2+)-sensitive nuclear signaling cascades for transcriptional up-regulation of cardiomyocyte hypertrophy.

• Authors: T G Valencia, L D Roberts, H Zeng, S R Grant
• Journal: Biochemical and biophysical research communications (Biochem Biophys Res Commun) Vol. 274 Issue 3 Pg. 803-10 (Aug 11 2000) ISSN: 0006-291X [Print] United States
• PMID: 10924357 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright 2000 Academic Press.

Chemical References

• Protein Synthesis Inhibitors
• Calcium-Calmodulin-Dependent Protein Kinase Type 2
• Calcium-Calmodulin-Dependent Protein Kinases
• Tetracycline

Topics

• Animals
• Calcium-Calmodulin-Dependent Protein Kinase Type 2
• Calcium-Calmodulin-Dependent Protein Kinases (physiology)
• Cell Size (physiology)
• Cells, Cultured
• Gene Expression Regulation, Enzymologic (drug effects, physiology)
• Mice
• Myocardium (metabolism, pathology)
• Protein Synthesis Inhibitors (pharmacology)
• Rats
• Rats, Sprague-Dawley
• Signal Transduction (drug effects)
• Tetracycline (pharmacology)