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Molecular and Cellular Biology, November 2000, p. 8397-8408, Vol. 20, No. 22
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Cdc13 Cooperates with the Yeast Ku Proteins and Stn1 To Regulate Telomerase Recruitment

Nathalie Grandin, Christelle Damon, and Michel Charbonneau*

Ecole Normale Supérieure, UMR CNRS/ENS 5665, Lyon 69364, France

Received 18 May 2000/Returned for modification 7 July 2000/Accepted 22 August 2000

The Saccharomyces cerevisiae CDC13 protein binds single-strand telomeric DNA. Here we report the isolation of new mutant alleles of CDC13 that confer either abnormal telomere lengthening or telomere shortening. This deregulation not only depended on telomerase (Est2/TLC1) and Est1, a direct regulator of telomerase, but also on the yeast Ku proteins, yKu70/Hdf1 and yKu80/Hdf2, that have been previously implicated in DNA repair and telomere maintenance. Expression of a Cdc13-yKu70 fusion protein resulted in telomere elongation, similar to that produced by a Cdc13-Est1 fusion, thus suggesting that yKu70 might promote Cdc13-mediated telomerase recruitment. We also demonstrate that Stn1 is an inhibitor of telomerase recruitment by Cdc13, based both on STN1 overexpression and Cdc13-Stn1 fusion experiments. We propose that accurate regulation of telomerase recruitment by Cdc13 results from a coordinated balance between positive control by yKu70 and negative control by Stn1. Our results represent the first evidence of a direct control of the telomerase-loading function of Cdc13 by a double-strand telomeric DNA-binding complex.

* Corresponding author. Mailing address: Ecole Normale Supérieure de Lyon, UMR CNRS 5665, 46, allée d'Italie, 69364 Lyon, France. Phone: (33) 472-72-81-70. Fax: (33) 472-72-80-80. E-mail: Michel.Charbonneau{at}ens-lyon.fr.

Molecular and Cellular Biology, November 2000, p. 8397-8408, Vol. 20, No. 22
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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