(Circulation. 1997;96:3943-3953.)
© 1997 American Heart Association, Inc.
Articles |
cis-Acting Sequences That Mediate Induction of ß-Myosin Heavy Chain Gene Expression During Left Ventricular Hypertrophy due to Aortic Constriction
From the Cardiovascular Division, Departments of Medicine and Cell Biology, Albert Einstein College of Medicine, Bronx, NY (K.H., S.J.L., R.N.K.), and the Department of Cell Biology, Parke-Davis Pharmaceutical Research Division, Warner Lambert Co, 2800 Plymouth Rd, Ann Arbor, Mich (S.M.J., B.E.M.).
Correspondence to Richard N. Kitsis, Departments of Medicine (Cardiology) and Cell Biology, Albert Einstein College of Medicine, 1300 Morris Park Ave, Bronx, NY 10461. E-mail kitsis{at}aecom.yu.edu
Background Marked alterations in the expression of specific genes occur during the development of cardiac hypertrophy in vivo. Little is known, however, about the cis-acting elements that mediate these changes in response to clinically relevant hypertrophic stimuli, such as hemodynamic overload, in intact adult animals.
Methods and Results The left ventricular expression
of a directly injected reporter gene driven by 3542 bp of rat
ß-myosin heavy chain (ß-MHC) promoter was increased 3.0-fold by
aortic constriction (P<.005), an increment similar to
the 3.2-fold increase in the level of the endogenous
ß-MHC mRNA in the same left ventricles. Subsequent analysis
identified a 107-bp ß-MHC promoter sequence (-303/-197) sufficient
to convert a heterologous neutral promoter to one that is
activated by aortic constriction. These sequences contain two
M-CAT elements, which have previously been demonstrated to mediate
inducible expression during 
1-adrenergic–stimulated
hypertrophy in cultured neonatal cardiac myocytes, and a
GATA element. Although simultaneous mutation of both M-CAT
elements markedly decreased the basal transcriptional activity of an
injected 333-bp ß-MHC promoter, it had no effect on aortic
constriction-stimulated transcription (3.5-fold increase,
P<.005 for both wild type and mutant). In contrast,
mutation of the GATA motif markedly attenuated aortic
constriction-stimulated transcription (1.6-fold, P=NS)
without affecting the basal transcriptional activity. This GATA site
can interact with in vitro translated GATA-4 and compete with an
established GATA site for GATA-4 binding activity in nuclear extracts
from aortic constricted hearts.
Conclusions Basal and aortic constriction-stimulated transcription of the ß-MHC gene is mediated, at least in part, through different mechanisms. A GATA element within ß-MHC sequences -303/-197 plays a role in the transcriptional activation of this gene by aortic constriction.
Key Words: genes • hypertrophy • signal transduction
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