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(Received for publication, April 24, 1996, and in revised form, September 3, 1996)
From the Laboratories of Biochemistry, Department of Animal
Biology, School of Veterinary Medicine, University of Pennsylvania,
Philadelphia, Pennsylvania 19104-6047
We have characterized the rat gene for
muscle-specific cytochrome oxidase VIII (COX VIII(H)) and mapped the
distal promoter region responsible for transcription activation in
C2C12 skeletal myocytes and H9C2 cardiomyocytes. In both cell types,
the promoter elements responding to the induced differentiation of
myocytes map to two E boxes, designated as E1 and E2 boxes with a core sequence of CAGCTG. Gel mobility shift analysis showed that both E1 and
E2 box motifs form complexes with nuclear extracts from H9C2
cardiomyocytes that were supershifted with monoclonal antibody to E2A
but not with antibody to myo-D. Extracts from induced and uninduced
H9C2 cardiomyocytes yielded different gel mobility patterns and also
different E2A antibody supershifts suggesting a difference in the
DNA-bound protein complexes cross-reacting with the E2A antibody.
Transcriptional activity of the promoter construct containing intact E
boxes was inhibited by coexpression with Id in differentiated H9C2
cardiomyocytes. Our results show the involvement of an E box binding
basic helix loop helix protein in the cardiac muscle-specific regulation of the COX VIII(H) promoter.
Volume 271, Number 47,
Issue of November 22, 1996
pp. 30281-30289
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
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