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Nucleic Acids Research 2005 33(8):2742-2750; doi:10.1093/nar/gki560
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Published online 12 May 2005

© The Author 2005. Published by Oxford University Press. All rights reserved
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Article

Chromatin assembly factor Asf1p-dependent occupancy of the SAS histone acetyltransferase complex at the silent mating-type locus HML{alpha}

Shigehiro Osada*, Mitsumasa Kurita, Jun-ichi Nishikawa and Tsutomu Nishihara

Laboratory of Environmental Biochemistry, Graduate School of Pharmaceutical Sciences, Osaka University 1-6 Yamada-Oka, Suita, Osaka 565-0871, Japan

*To whom correspondence should be addressed. Tel: +81 6 6879 8242; Fax: +81 6 6879 8244; Email: osada{at}phs.osaka-u.ac.jp

Received January 28, 2005. Revised April 20, 2005. Accepted April 20, 2005.

Transcriptional repression of the silent mating-type loci HML{alpha} and HMRa in Saccharomyces cerevisiae is regulated by chromatin structure. Sas2p is a catalytic subunit of the SAS histone acetyltransferase (HAT) complex. Although many HATs seem to relieve chromosomal repression to facilitate transcriptional activation, sas mutant phenotypes include loss of SIR1-dependent silencing of HML{alpha}. To gain insight into the mechanism of the SAS complex mediated silencing at HML{alpha}, we investigated the expression and chromatin structure of the {alpha}2 gene in the HML{alpha} locus. We found that deletion of SAS2 in combination with a null allele of SIR1 changed the chromatin structure of the precisely positioned nucleosome, which includes the mRNA start site of the {alpha}2 gene and derepressed {alpha}2 transcription. The Sas2p HAT domain was required for this silencing. Furthermore, chromatin immunoprecipitation analysis revealed that the SAS complex was associated with the HML{alpha} locus, and ASF1 (which encodes chromatin assembly factor Asf1p), but not SIR1 and SIR2, was necessary for this localization. These data suggest that the HAT activity and ASF1-dependent localization of the SAS complex are required for SIR1-dependent HML{alpha} silencing.


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