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Originally published In Press as doi:10.1074/jbc.M004294200 on June 23, 2000

J. Biol. Chem., Vol. 275, Issue 37, 28363-28370, September 15, 2000
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DNA Binding Specificity Studies of Four ETS Proteins Support an Indirect Read-out Mechanism of Protein-DNA Recognition*,

Blair R. Szymczyna and Cheryl H. ArrowsmithDagger

From the Ontario Cancer Institute and Department of Medical Biophysics, University of Toronto, Toronto, Ontario M5G 2M9, Canada

Members of the ETS family of transcription factors are involved in several developmental and physiological processes, and, when overexpressed or misexpressed, can contribute to a variety of cancers. Each family member has a conserved DNA-binding domain that recognizes DNA sequences containing a G-G-A trinucleotide. Discrimination between potential ETS-binding sites appears to be governed by both the nucleotides flanking the G-G-A sequence and protein-protein interactions. We have used an adaptation of the "length-encoded multiplex" approach (Desjarlais, J. R., and Berg, J. M. (1994) Proc. Natl. Acad. Sci. U. S. A. 91, 11099-11103) to define DNA binding specificities for four ETS proteins: Fli-1, SAP-1, PU.1, and TEL. Our results support a model in which cooperative effects among neighboring bases flanking the central G-G-A site contribute to the formation of stable ETS/DNA complexes. These results are consistent with a mechanism for specific DNA binding that is partially governed by an indirect read-out of the DNA sequence, in which a sequence-specific DNA conformation is sensed or induced.

* This work was supported by the Medical Research Council of Canada.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.

The on-line version of this article (available at http://www.jbc.com) contains supplementary tables.

Dagger To whom correspondence should be addressed: Ontario Cancer Inst. and Dept. of Medical Biophysics, University of Toronto, 610 University Ave., Toronto, ON M5G 2M9, Canada. Tel.: 416-946-2017; Fax: 416-946-6529; E-mail: carrow@uhnres.utoronto.ca.

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