原创力文档regulation of budding yeast mating-type switching donor preference by the fha domain of fkh1监管出芽酵母接合型捐助偏好fkh1 fha域的转换.pdfVIP
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regulation of budding yeast mating-type switching donor preference by the fha domain of fkh1监管出芽酵母接合型捐助偏好fkh1 fha域的转换
Regulation of Budding Yeast Mating-Type Switching
Donor Preference by the FHA Domain of Fkh1
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Jin Li , Eric Coıc , Kihoon Lee, Cheng-Sheng Lee, Jung-Ae Kim , Qiuqin Wu, James E. Haber*
Department of Biology and Rosenstiel Center, Brandeis University, Waltham, Massachusetts, United States of America
Abstract
During Saccharomyces cerevisiae mating-type switching, an HO endonuclease-induced double-strand break (DSB) at MAT is
repaired by recombining with one of two donors, HMLa or HMRa, located at opposite ends of chromosome III. MATa cells
preferentially recombine with HMLa; this decision depends on the Recombination Enhancer (RE), located about 17 kb to the
right of HML. In MATa cells, HML is rarely used and RE is bound by the MATa2-Mcm1 corepressor, which prevents the
binding of other proteins to RE. In contrast, in MATa cells, RE is bound by multiple copies of Fkh1 and a single copy of Swi4/
Swi6. We report here that, when RE is replaced with four LexA operators in MATa cells, 95% of cells use HMR for repair, but
expression of a LexA-Fkh1 fusion protein strongly increases HML usage. A LexA-Fkh1 truncation, containing only Fkh1’s
phosphothreonine-binding FHA domain, restores HML usage to 90%. A LexA-FHA-R80A mutant lacking phosphothreonine
binding fails to increase HML usage. The LexA-FHA fusion protein associates with chromatin in a 10-kb interval surrounding
the HO cleavage site at MAT, but only after DSB induction. This association occurs even in a donorless strain lacking HML. We
propose that the FHA domain of Fkh1 regulates donor preference by physically interacting with phosphorylated threonine
residues created on proteins bound near the DSB, thus
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