Contraction accelerates myosin heavy chain synthesis rates in adult cardiocytes by an increase in the rate of translational initiation.
The purpose of this study was to determine the mechanism by which contraction acutely accelerates the synthesis rate of the contractile protein myosin heavy chain (MHC). Laminin-adherent adult feline cardiocytes were maintained in a serum-free medium and induced to contract at 1 Hz via electrical field stimulation. Electrical stimulation of contraction accelerated rates of MHC synthesis 28%, p < 0.05 by 4 h as determined by incorporation of [3H]phenylalanine into MHC. MHC mRNA expression as measured by RNase protection was unchanged after 4 h of electrical stimulation. MHC mRNA levels in messenger ribonucleoprotein complexes and translating polysomes were examined by sucrose gradient fractionation. The relative percentage of polysomebound MHC mRNA was equal at 47% in both electrically stimulated and control cardiocytes. However, electrical stimulation of contraction resulted in a reproducible shift of MHC mRNA from smaller polysomes into larger polysomes, indicating an increased rate of initiation. This shift resulted in significant increases in MHC mRNA levels in the fractions containing the larger polysomes of electrically stimulated cardiocytes as compared with nonstimulated controls. These data indicate that the rate of MHC synthesis is accelerated in contracting cardiocytes via an increase in translational efficiency.
Ivester
C T
CT
Department of Medicine, Gazes Cardiac Research Institute, Charleston, South Carolina, USA.
Tuxworth
W J
WJ
Cooper
G
G
McDermott
P J
PJ
D000818:Animals; D002415:Cats; D002478:Cells, Cultured; D015342:DNA Probes; D004558:Electric Stimulation / Q000295:instrumentation / Q000379:methods; D005786:Gene Expression Regulation; D006321:Heart / Q000502:physiology*; D007700:Kinetics; D009200:Myocardial Contraction; D009206:Myocardium / Q000378:metabolism*; D009218:Myosins / Q000096:biosynthesis*; D010649:Phenylalanine / Q000378:metabolism; D011132:Polyribosomes / Q000378:metabolism; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D012270:Ribosomes / Q000378:metabolism; D014316:Tritium
D015342:DNA Probes; D012333:RNA, Messenger; D014316:Tritium; D010649:Phenylalanine; D009218:Myosins