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J Biol Chem, Vol. 273, Issue 28, 17865-17870, July 10, 1998
From the Cells were transfected with luciferase reporter
genes, under the control of promoters derived from either the farnesyl
diphosphate (FPP) synthase, 3-hydroxy-3-methylglutaryl coenzyme A
(HMG-CoA) synthase, HMG-CoA reductase, or low density lipoprotein
receptor genes. The increase in luciferase activity that occurred when cells were either incubated in sterol-depleted medium or cotransfected with a cDNA encoding sterol regulatory element-binding protein (SREBP)-1a was prevented by coexpression of wild-type E1A or a Gal4-CBP
(1-451) fusion protein. The inhibitory effect of E1A was overcome by
coexpression of CBP. The increase in reporter gene activity noted above
was not affected when the cells were cotransfected with cDNAs that
encoded either a mutant E1A that is unable to interact with the
transcriptional activator CBP or Gal4-CBP fusion proteins encoding
separate fragments of CBP, which span the remainder of the CBP
molecule. A preformed SREBP-1a:[32P]DNA complex
bound specifically to membrane-immobilized GST-CBP fusion proteins that
contained amino-terminal portions of CBP.
In order to investigate the role of CBP in the regulation of endogenous
genes, we isolated stable transformants that express Gal4-CBP(1-451)
in response to added doxycycline. Induction of endogenous FPP synthase
and HMG-CoA synthase mRNAs, in response to cellular cholesterol
depletion, was prevented when cells expressed Gal4-CBP(1-451).
We conclude that when cells are incubated in the absence of sterols,
the transcriptional activation of the HMG-CoA synthase, HMG-CoA
reductase, FPP synthase, and low density lipoprotein receptor genes is
dependent on a specific interaction between SREBP, which is bound to
the promoter DNA, and the amino-terminal domain (amino acids 1-451) of
CBP.
CBP Is Required for Sterol-regulated and Sterol Regulatory
Element-binding Protein-regulated Transcription
and¶
Departments of Biological Chemistry and
Medicine and the ¶ Molecular Biology Institute, UCLA, Los
Angeles, California 90095
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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