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Vol. 273, Issue 3, 1349-1356, January 16, 1998
From the Department of Molecular Biology and Biochemistry,
University of California, Irvine, California 92697-3900
3-Hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA)
synthase, a key regulatory enzyme in the pathway for endogenous
cholesterol synthesis, is a target for negative feedback regulation by
cholesterol. The promoter for HMG-CoA synthase contains two
binding sites for the sterol regulatory element-binding proteins
(SREBPs). When cellular sterol levels are low, the SREBPs are
released from the endoplasmic reticulum membrane, allowing them to
translocate to the nucleus and activate SREBP target genes. In all
SREBP-regulated promoters studied to date, additional co-regulatory
transcription factors are required. In the HMG-CoA synthase promoter
there are several potential co-regulatory transcription factor binding
sites, including an inverted CCAAT box. A similar element has been
shown to function with SREBP to mediate sterol regulation of another gene involved in cholesterol metabolism, farnesyl diphosphate synthase. Here, we show that CCAAT binding factor/nuclear factor Y (CBF/NF-Y) binding to the CCAAT box is required for sterol-regulated transcription of HMG-CoA synthase. The SREBP sites and the inverted CCAAT box are normally separated by 17 base pairs, and we show that
increasing this distance results in a decrease in the level of
transcriptional regulation by sterols. Furthermore, we provide evidence
that there is a direct interaction between CBF/NF-Y and the basic
helix-loop-helix-zipper region of SREBP. Interestingly, this
interaction does not occur efficiently with any of the isolated subunits and appears to require all three nonidentical CBF/NF-Y subunits in a preassembled complex. Since CBF/NF-Y only binds to DNA
when all three subunits are in a complex, this would prevent SREBP from forming nonproductive associations with the individual subunits.
Sterol Regulation of 3-Hydroxy-3-Methylglutaryl-coenzyme A
Synthase Gene through a Direct Interaction Between Sterol Regulatory
Element Binding Protein and the Trimeric CCAAT-binding
Factor/Nuclear Factor Y
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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