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Regulation of budding yeast mating-type switching donor preference by the FHA domain of Fkh1.
PLoS Genet. 2012; 8(4):e1002630.PG

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, HMLα or HMRa, located at opposite ends of chromosome III. MATa cells preferentially recombine with HMLα; this decision depends on the Recombination Enhancer (RE), located about 17 kb to the right of HML. In MATα cells, HML is rarely used and RE is bound by the MATα2-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 (γ-H2AX) around the DSB but can also promote γ-H2AX spreading around the RE region.

Authors+Show Affiliations

Department of Biology and Rosenstiel Center, Brandeis University, Waltham, Massachusetts, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural

Language

eng

PubMed ID

22496671

Citation

Li, Jin, et al. "Regulation of Budding Yeast Mating-type Switching Donor Preference By the FHA Domain of Fkh1." PLoS Genetics, vol. 8, no. 4, 2012, pp. e1002630.
Li J, Coïc E, Lee K, et al. Regulation of budding yeast mating-type switching donor preference by the FHA domain of Fkh1. PLoS Genet. 2012;8(4):e1002630.
Li, J., Coïc, E., Lee, K., Lee, C. S., Kim, J. A., Wu, Q., & Haber, J. E. (2012). Regulation of budding yeast mating-type switching donor preference by the FHA domain of Fkh1. PLoS Genetics, 8(4), e1002630. https://doi.org/10.1371/journal.pgen.1002630
Li J, et al. Regulation of Budding Yeast Mating-type Switching Donor Preference By the FHA Domain of Fkh1. PLoS Genet. 2012;8(4):e1002630. PubMed PMID: 22496671.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Regulation of budding yeast mating-type switching donor preference by the FHA domain of Fkh1. AU - Li,Jin, AU - Coïc,Eric, AU - Lee,Kihoon, AU - Lee,Cheng-Sheng, AU - Kim,Jung-Ae, AU - Wu,Qiuqin, AU - Haber,James E, Y1 - 2012/04/05/ PY - 2011/12/14/received PY - 2012/02/17/accepted PY - 2012/4/13/entrez PY - 2012/4/13/pubmed PY - 2012/9/27/medline SP - e1002630 EP - e1002630 JF - PLoS genetics JO - PLoS Genet VL - 8 IS - 4 N2 - 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, HMLα or HMRa, located at opposite ends of chromosome III. MATa cells preferentially recombine with HMLα; this decision depends on the Recombination Enhancer (RE), located about 17 kb to the right of HML. In MATα cells, HML is rarely used and RE is bound by the MATα2-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 (γ-H2AX) around the DSB but can also promote γ-H2AX spreading around the RE region. SN - 1553-7404 UR - https://www.unboundmedicine.com/medline/citation/22496671/Regulation_of_budding_yeast_mating_type_switching_donor_preference_by_the_FHA_domain_of_Fkh1_ L2 - https://dx.plos.org/10.1371/journal.pgen.1002630 DB - PRIME DP - Unbound Medicine ER -