Repressive Effect of Sp1 on the C/EBPalpha  Gene Promoter: Role in Adipocyte Differentiation

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Molecular and Cellular Biology, July 1999, p. 4855-4865, Vol. 19, No. 7
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Repressive Effect of Sp1 on the C/EBP $alpha$ Gene Promoter: Role in Adipocyte Differentiation

Qi-Qun Tang, Man-Shiow Jiang, and M. Daniel Lane^*

Department of Biological Chemistry, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205

Received 22 October 1998/Returned for modification 6 January 1999/Accepted 21 April 1999

Expression of C/EBP $alpha$ is required for differentiation of 3T3-L1 preadipocytes into adipocytes. Previous investigations indicatedthat transcription of the C/EBP $alpha$ gene is sequentially activatedduring differentiation, initially by C/EBP $beta$ and C/EBP $delta$ and laterby C/EBP $alpha$ (autoactivation). These events are mediated by a C/EBPregulatory element in the promoter of the C/EBP $alpha$ gene. This articlepresents evidence that members of the Sp family, notably Sp1,act repressively on the C/EBP $alpha$ promoter prior to the inductionof differentiation. Sp1 was shown to bind to a GC box at the 5'end of the C/EBP regulatory element in the C/EBP $alpha$ promoter and,in so doing, to competitively prevent binding to and transactivationof the promoter by the C/EBPs. One of the differentiation inducersmethylisobutylxanthine (a cAMP phosphodiesterase inhibitor) orForskolin, both of which increase the cellular cAMP level, causesdown-regulation of Sp1. This decrease in Sp1 level early in thedifferentiation program appears to facilitate access of C/EBP $beta$ and/or C/EBP $delta$ to the C/EBP regulatory element and, thereby, derepressionof the C/EBP $alpha$ gene.

^* Corresponding author. Mailing address: Department of Biological Chemistry, The Johns Hopkins University School of Medicine,725 N. Wolfe St., Baltimore, MD 21205. Phone: (410) 955-3554.Fax: (410) 955-0903. E-mail: dlane{at}jhmi.edu.

Molecular and Cellular Biology, July 1999, p. 4855-4865, Vol. 19, No. 7
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

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Repressive Effect of Sp1 on the C/EBP Gene Promoter: Role in Adipocyte Differentiation

Repressive Effect of Sp1 on the C/EBP $alpha$ Gene Promoter: Role in Adipocyte Differentiation