Creatine kinase is an alpha myosin heavy chain 3'UTR mRNA binding protein.
Altered cardiac workload regulates the translation and localization of the alpha myosin heavy chain (alphaMyHC) messenger RNA through the 3' untranslated region (UTR) by protein-RNA interactions. We used the alphaMyHC 3'UTR from neonatal rat heart tissue in a gel shift analysis to find RNA binding proteins. One was identified by microsequencing as creatine kinase, brain form B (CKBB). The affinity of its binding interaction was evaluated using sense and antisense alphaMyHC 3'UTR and 3'UTR probes from myosin isoforms of 2B and 2X skeletal muscle. Removal of calcium by the chelating agent EGTA had a potentiating effect on the formation of the CKBB/alphaMyHC 3'UTR complex in vitro . Varying the concentration of ATP (0.1-1 mM) also enhanced this interaction, suggesting that autophosphorylation of CKBB is taking place. Our novel finding that CKBB, an energy transduction enzyme, binds to the RNA of the 3'UTR of the faster ATP consuming alphaMyHC suggests a possible regulatory linkage between the metabolic state of the cell and myosin isoform expression.
Vracar-Grabar
Marina
M
Department of Physiology and Biophysics (M/C 901), University of Illinois at Chicago, 835 S. Wolcott Avenue, Chicago, IL 60612-1342, USA.
Russell
Brenda
B
D020413:3' Untranslated Regions / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000378:metabolism; D003001:Cloning, Molecular; D003402:Creatine Kinase / Q000378:metabolism*; D004533:Egtazic Acid / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional; D007545:Isoproterenol / Q000494:pharmacology; D009206:Myocardium / Q000201:enzymology*; D018995:Myosin Heavy Chains / Q000378:metabolism*; D010766:Phosphorylation / Q000187:drug effects; D011485:Protein Binding / Q000187:drug effects; D012333:RNA, Messenger / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D024723:Ventricular Myosins / Q000378:metabolism*; D014700:Verapamil / Q000494:pharmacology
D020413:3' Untranslated Regions; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D004533:Egtazic Acid; D000255:Adenosine Triphosphate; D014700:Verapamil; D003402:Creatine Kinase; D024723:Ventricular Myosins; D018995:Myosin Heavy Chains; D007545:Isoproterenol; D002118:Calcium