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We have studied the effect of several myogenic regulatory
factors on the activity of the promoter for a mouse gene encoding a
skeletal myosin heavy chain (MyHC) expressed in adult (type IIB) muscle
fibers. Co-transfection of myogenic factors is necessary for activity
of the IIB promoter in mouse C2 myotubes in culture but not in quail
myotubes in culture. Although this promoter contains one E-box within
the first 192 base pairs upstream of the transcriptional start site,
mutations in this motif demonstrate that it is not required for the
transactivation effect of the myogenic factors. Analysis of other
mutants suggests that the MEF2 and MHox DNA-binding factor binds to an
evolutionarily conserved AT-rich motif. In addition, the IIB promoter
appears to require the conserved TATA motif (CTATAAAAG) in order to be
activated by the AT-rich sequences. The IIB promoter constructs produce
RNA transcripts which begin at the natural site of transcriptional
initiation in quail myotubes and in mouse C2 myotubes after
co-transfection with myogenic factors; a second, minor, start site is
also used in the co-transfected C2 myotubes. Results obtained after
transfection of the mouse IIB promoter constructs in quail myotube
cultures suggest that the overexpression of myogenic factors in C2
cultures does not result in an environment in which the control of IIB
MyHC promoter activity is aberrant. Therefore, either the myogenic
factors themselves, or other proteins induced by them, seem to interact
directly with the basal transcription machinery to allow
muscle-specific gene expression.
Volume 270,
Number 26,
Issue of June 30, pp. 15664-15670, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
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