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(Received for publication, May 7, 1996, and in revised form, July 31, 1996)
From the Department of Biochemistry and Molecular Biology and the
Norris Comprehensive Cancer Center, University of Southern California
School of Medicine, Los Angeles, California 90033-0800
Transcription of the gene encoding GRP78/BiP, a
calcium-binding molecular chaperone localized in the endoplasmic
reticulum, is induced in mammalian cells through gradual depletion of
the intracellular calcium stores. The multimeric CCAAT binding factor, CBF/NF-Y, binds to the most proximal CCAAT regulatory element (C1) of
the grp78 promoter required for both basal level expression and stress response. Using an in vitro transcription
system, we show through factor competition and immunodepletion that the
grp78 C1-mediated enhancement of transcription requires
primarily CBF. Correlating with the previous observation that CBF
binding to the 78C1 site is enhanced by EGTA and EDTA, these divalent
cation chelators specifically stimulate 78C1-directed transcription. In
contrast, increasing amounts of calcium ions are inhibitory. These
results provide evidence that CBF is functionally important in
transactivating the grp78 C1 transcriptional activity, and suggest a possible mechanism by which grp78 transcription
is stimulated by calcium depletion. We further discovered that in
addition to binding CBF, both the 78C1 element and the CBF binding site
of the
Volume 271, Number 46,
Issue of November 15, 1996
pp. 28995-29002
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
2(I) collagen promoter interact weakly with the
multifunctional transcription factor YY1. Our studies show that the
binding sites for CBF and YY1 are distinct for the two promoter sites,
suggesting that YY1 and other interacting factors could exert
differential effects on individual promoters bearing the same CBF
site.
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