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Originally published In Press as doi:10.1074/jbc.M000931200 on July 17, 2000

J. Biol. Chem., Vol. 275, Issue 38, 29701-29708, September 22, 2000
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Characterization of the Activation Domains of AP-2 Family Transcription Factors*

Sharad WankhadeDagger , Yihong YuDagger , Justin WeinbergDagger , Michael A. Tainsky§, and Perry KannanDagger

From the Dagger  Rammelkamp Center for Education and Research, MetroHealth Campus, Case Western Reserve University, Cleveland, Ohio 44109 and § Program in Molecular Biology and Genetics, Karmanos Cancer Institute, Wayne State University, Detroit, Michigan 48201

Despite sequence variation, all AP-2 isotypes are capable of activating transcription, which indicates a functional conservation. We used this property to gain a unique insight into the structure and function of the activation motifs of AP-2 family transcription factors. We have precisely localized the activation motif of human AP-2alpha to amino acids 52-108. Our experiments indicate that similar sequence of amino acids in all AP-2 isotypes except Drosophila AP-2alpha harbor their activation motifs. Within this sequence, fewer than 36 residues are critical for transcription activation. Our comparison studies and site-directed mutagenic analyses show that these critical amino acids are strategically placed within this sequence. These residues are interspersed with nonessential and influential residues that vary in composition and length, indicating a structural flexibility. The Drosophila AP-2alpha has its partly conserved activation motif in an extended region about twice the length of other AP-2 isotypes. Our results reveal essential elements of the amino acid composition of activators in general and shed new light on the mechanism of transcription activation.

* This work was supported by Grants CA67036 and CA84278 (to P. K.) and CA53475 (to M. A. T.) from the NCI, National Institutes of Health.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed: Rammelkamp Center for Education and Research, MetroHealth Campus, Case Western Reserve University, 2500 MetroHealth Dr., Cleveland, OH 44109. Tel.: 216-778-1156; Fax: 216-778-2770; E-mail: pkannan@research.metrohealth.org.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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