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J. Biol. Chem., Vol. 278, Issue 31, 29327-29335, August 1, 2003

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Flexible DNA Binding of the BTB/POZ-domain Protein FBI-1^*

Frank Pessler   || and Nouria Hernandez  ¶

From the Cold Spring Harbor Laboratory and ^¶Howard Hughes Medical Institute, Cold Spring Harbor, New York 11724

POZ-domain transcription factors are characterized by the presence of a protein-protein interaction domain called the POZ or BTB domain at their N terminus and zinc fingers at their C terminus. Despite the large number of POZ-domain transcription factors that have been identified to date and the significant insights that have been gained into their cellular functions, relatively little is known about their DNA binding properties. FBI-1 is a BTB/POZ-domain protein that has been shown to modulate HIV-1 Tat trans-activation and to repress transcription of some cellular genes. We have used various viral and cellular FBI-1 binding sites to characterize the interaction of a POZ-domain protein with DNA in detail. We find that FBI-1 binds to inverted sequence repeats downstream of the HIV-1 transcription start site. Remarkably, it binds efficiently to probes carrying these repeats in various orientations and spacings with no particular rotational alignment, indicating that its interaction with DNA is highly flexible. Indeed, FBI-1 binding sites in the adenovirus 2 major late promoter, the c-fos gene, and the c-myc P1 and P2 promoters reveal variously spaced direct, inverted, and everted sequence repeats with the consensus sequence G(A/G)GGG(T/C)(C/T)(T/C)(C/T) for each repeat.

Received for publication, March 24, 2003 , and in revised form, May 14, 2003.

^* 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.

Present address: Div. of Rheumatology, Children's Hospital of Philadelphia, 3516 Civic Center Blvd./1102 ARC, Philadelphia, PA 19104.

^|| To whom correspondence should be addressed. Tel.: 215-590-7180; Fax: 215-590-1258; E-mail: pessler{at}email.chop.edu.

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