Regulation of budding yeast mating-type switching donor preference by the FHA domain of Fkh1.

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Show simple item record Li, Jin Coïc, Eric Lee, Kihoon Lee, Cheng-Sheng Kim, Jung-Ae Wu, Qiuqin Haber, James E 2019-01-29T18:18:28Z 2019-01-29T18:18:28Z 2012
dc.identifier.issn 1553-7390
dc.identifier.issn 1553-7404
dc.description.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, HML ± or HMRa, located at opposite ends of chromosome III. MATa cells preferentially recombine with HML ±; this decision depends on the Recombination Enhancer (RE), located about 17 kb to the right of HML. In MAT ± cells, HML is rarely used and RE is bound by the MAT ±2-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 positioning HML close to the DSB at MAT. Donor preference is independent of Mec1/ATR and Tel1/ATM checkpoint protein kinases but partially depends on casein kinase II. RE stimulates the strand invasion step of interchromosomal recombination even for non-MAT sequences. We also find that when RE binds to the region near the DSB at MATa then Mec1 and Tel1 checkpoint kinases are not only able to phosphorylate histone H2A ( _-H2AX) around the DSB but can also promote _-H2AX spreading around the RE region.
dc.format.extent 1 file
dc.language English
dc.language.iso eng
dc.publisher Public Library of Science
dc.rights Creative Commons Attribution 4.0 International License
dc.subject mating type
dc.subject cell cycle checkpoint
dc.subject histone
dc.subject chromatin
dc.subject phosphorylation
dc.subject fusion protein
dc.subject protein kinase a
dc.subject dna repair
dc.subject dna binding protein
dc.subject recombinant dna
dc.subject polymerase chain reaction
dc.subject molecular biology
dc.subject biochemistry
dc.subject genetics
dc.subject biology
dc.title Regulation of budding yeast mating-type switching donor preference by the FHA domain of Fkh1.
dc.type Article
dc.contributor.department Department of Biology
dc.relation.journal PLoS Genetics
dc.identifier.pmid 22496671
dc.identifier.pmcid PMC3320585
dc.description.esploro yes

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