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J. Biol. Chem., Vol. 281, Issue 40, 30122-30131, October 6, 2006

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Characterization of Distant Enhancers and Promoters in the Albumin--Fetoprotein Locus during Active and Silenced Expression^*

From the Department of Pathology, Albert Einstein College of Medicine, Bronx, New York 10461

The albumin and -fetoprotein genes are adjacent and express closely related serum proteins. Both genes are strongly expressed in fetal liver, primarily through activation by distant enhancers, but the AFP gene selectively undergoes developmental silencing. We used chromatin immunoprecipitation to study enhancers and promoters during active and silenced gene expression. In adult phenotype cells, the silenced AFP gene was actively repressed at the promoter and two proximal enhancers, characterized by the absence of coactivators and acetylated histone 4, and the presence of corepressors and K9-methylated histone 3. Specific transcription factors, TBP, and RNA polymerase II were all detected on both active and silenced genes, indicating that both states were actively regulated. Surprisingly, promoter-specific factors were also detected on enhancers, especially with reduced chromatin shearing. Under these conditions, an enhancer-specific factor was also detected on the albumin promoter. Association of promoter- and enhancer-specific factors was confirmed by sequential immunoprecipitation. Because no binding was detected on intervening segments, these promoter-enhancer associations suggest looping.

Received for publication, April 11, 2006 , and in revised form, July 5, 2006.

^* This work was supported by Grants CA68440 and CA76354 from the National Institutes of Health. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ To whom correspondence should be addressed: Dept. of Pathology, Albert Einstein College of Medicine, 1300 Morris Park Ave., Bronx, NY 10461-1975. Tel.: 718-430-3422; Fax: 718-430-3483; E-mail: locker{at}aecom.yu.edu.

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