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Genetics, Vol. 150, 613-632, October 1998, Copyright © 1998

Identification of High-Copy Disruptors of Telomeric Silencing in Saccharomyces cerevisiae

Miriam S. Singera, Alon Kahanaa,b, Alexander J. Wolfb, Lia L. Meisingera, Suzanne E. Petersonb, Colin Goggina, Maureen Mahowalda, and Daniel E. Gottschlingb
a Department of Molecular Genetics and Cell Biology, The University of Chicago, Chicago, Illinois 60637
b Division of Basic Sciences, Fred Hutchinson Cancer Research Center, Seattle, Washington 98109

Corresponding author: Daniel E. Gottschling, Division of Basic Sciences, Mail-Stop A3-025, Fred Hutchinson Cancer Research Center, 1100 Fairview Ave. North, P.O. Box 19024, Seattle, WA 98109-1024., dgottsch{at}fhcrc.org (E-mail).

Communicating editor: F. WINSTON

The ends of chromosomes in Saccharomyces cerevisiae initiate a repressive chromatin structure that spreads internally and inhibits the transcription of nearby genes, a phenomenon termed telomeric silencing. To investigate the molecular basis of this process, we carried out a genetic screen to identify genes whose overexpression disrupts telomeric silencing. We thus isolated 10 DOT genes (disruptor of telomeric silencing). Among these were genes encoding chromatin component Sir4p, DNA helicase Dna2p, ribosomal protein L32, and two proteins of unknown function, Asf1p and Ifh1p. The collection also included genes that had not previously been identified: DOT1, DOT4, DOT5, DOT6, and TLC1, which encodes the RNA template component of telomerase. With the exception of TLC1, all these genes, particularly DOT1 and DOT4, also reduced silencing at other repressed loci (HM loci and rDNA) when overexpressed. Moreover, deletion of the latter two genes weakened silencing as well, suggesting that DOT1 and DOT4 normally play important roles in gene repression. DOT1 deletion also affected telomere tract length. The function of Dot1p is not known. The sequence of Dot4p suggests that it is a ubiquitin-processing protease. Taken together, the DOT genes include both components and regulators of silent chromatin.




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