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Molecular and Cellular Biology, October 2001, p. 6606-6614, Vol. 21, No. 19
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.19.6606-6614.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Targeting of the Yeast Ty5 Retrotransposon to Silent Chromatin Is Mediated by Interactions between Integrase and Sir4p†

Weiwu Xie,1 Xiaowu Gai,1 Yunxia Zhu,1 David C. Zappulla,2 Rolf Sternglanz,2 and Daniel F. Voytas1,*

Department of Zoology and Genetics, Iowa State University, Ames, Iowa 50011-3260,1 and Department of Biochemistry and Cell Biology, State University of New York, Stony Brook, New York 11794-52152

Received 12 March 2001/Returned for modification 8 May 2001/Accepted 30 June 2001

The Ty5 retrotransposons of Saccharomyces cerevisiae integrate preferentially into regions of silent chromatin at the telomeres and silent mating loci (HMR and HML). We define a Ty5-encoded targeting domain that spans 6 amino acid residues near the C terminus of integrase (LXSSXP). The targeting domain establishes silent chromatin when it is tethered to a weakened HMR-E silencer, and it disrupts telomeric silencing when it is overexpressed. As determined by both yeast two-hybrid and in vitro binding assays, the targeting domain interacts with the C terminus of Sir4p, a structural component of silent chromatin. This interaction is abrogated by mutations in the targeting domain that disrupt integration into silent chromatin, suggesting that recognition of Sir4p by the targeting domain is the primary determinant in Ty5 target specificity.

* Corresponding author. Mailing address: 2208 Molecular Biology Bldg., Iowa State University, Ames, IA 50011. Phone: (515) 294-1963. Fax: (515) 294-7155. E-mail: voytas{at}iastate.edu.

dagger This is Journal Paper J-19154 of the Iowa Agriculture and Home Economics Experiment Station, Ames, project 3383.

Molecular and Cellular Biology, October 2001, p. 6606-6614, Vol. 21, No. 19
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.19.6606-6614.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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