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J. Biol. Chem., Vol. 279, Issue 49, 50764-50772, December 3, 2004

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Taz1 Binding to a Fission Yeast Model Telomere

FORMATION OF TELOMERIC LOOPS AND HIGHER ORDER STRUCTURES^*

Lubomir Tomaska, Smaranda Willcox, Judita Slezakova, Jozef Nosek¶, and Jack D. Griffith||

From the Departments of Genetics and ^¶Biochemistry, Comenius University, Faculty of Natural Sciences, Mlynska dolina, 842 15 Bratislava, Slovakia and the Lineberger Comprehensive Cancer Center and Department of Microbiology and Immunology, University of North Carolina, Chapel Hill, North Carolina 27599

Similar to its human homologues TRF1 and TRF2, fission yeast Taz1 protein is a component of telomeric chromatin regulating proper telomere maintenance. As mammalian TRF1 and TRF2 proteins have been shown to directly bind telomeric DNA to form protein arrays and looped structures, termed t-loops, the ability of Taz1p to act on fission yeast telomeric DNA in similar ways was examined using purified protein and model DNA templates. When incubated with Taz1p, model telomeres containing 3' single-stranded telomeric overhangs formed t-loops at a frequency approaching 13%. Termini with blunt ends and non-telomeric overhangs were deficient in t-loop formation. In addition, we observed arrays of multiple Taz1p molecules bound to the telomeric regions, resembling the pattern of TRF1 binding. The presence of t-loops larger than the telomeric tract, a high frequency of end-bound DNAs and a donut shape of the Taz1p complex suggest that Taz1p binds the 3' overhang then extrudes a loop that grows in size as the donut slides along the duplex DNA. Based on these in vitro results we discuss possible general implications for fission yeast telomere dynamics.

Received for publication, August 25, 2004

^* This work was supported in part by Fogarty International Research Collaboration Award TW05654-01, Howard Hughes Medical Institute Grant 55000327, Slovak agency VEGA Grants 1/9153/02 and 1/0006/03, and Comenius University Grant UK/131/2002. 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.

^|| Supported by an Ellison Senior Scholar Award and National Institutes of Health Grants GM31819 and CA19014. To whom correspondence should be addressed: Lineberger Comprehensive Cancer Center, University of North Carolina, Mason Farm Road CB7095, Chapel Hill NC 27599-7295. Tel.: 919-966-2151; Fax: 919-966-3015; E-mail: jdg{at}med.unc.edu.

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