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.
Vyšlo v časopise: Regulation of Budding Yeast Mating-Type Switching Donor Preference by the FHA Domain of Fkh1. PLoS Genet 8(4): e32767. doi:10.1371/journal.pgen.1002630 Kategorie: Research Article prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1002630
1. HaberJE 1998 Mating-type gene switching in Saccharomyces cerevisiae. Annu Rev Genet 32 561 599
2. HaberJE 2012 Mating-type genes and MAT switching in Saccharomyces cerevisiae. Yeast Book Genetics Society of America In press
3. KlarAJHicksJBStrathernJN 1982 Directionality of yeast mating-type interconversion. Cell 28 551 561
4. LooSRineJ 1994 Silencers and domains of generalized repression. Science 264 1768 1771
5. RavindraAWeissKSimpsonRT 1999 High-resolution structural analysis of chromatin at specific loci: Saccharomyces cerevisiae silent mating-type locus HMRa. Mol Cell Biol 19 7944 7950
6. WeissKSimpsonRT 1998 High-resolution structural analysis of chromatin at specific loci: Saccharomyces cerevisiae silent mating type locus HMLalpha. Mol Cell Biol 18 5392 5403
7. WeilerKSBroachJR 1992 Donor locus selection during Saccharomyces cerevisiae mating type interconversion responds to distant regulatory signals. Genetics 132 929 942
8. WuXHaberJE 1995 MATa donor preference in yeast mating-type switching: activation of a large chromosomal region for recombination. Genes Dev 9 1922 1932
9. WuXMooreJKHaberJE 1996 Mechanism of MAT alpha donor preference during mating-type switching of Saccharomyces cerevisiae. Mol Cell Biol 16 657 668
10. WuCWeissKYangCHarrisMATyeBK 1998 Mcm1 regulates donor preference controlled by the recombination enhancer in Saccharomyces mating-type switching. Genes Dev 12 1726 1737
11. WuXHaberJE 1996 A 700 bp cis-acting region controls mating-type dependent recombination along the entire left arm of yeast chromosome III. Cell 87 277 285
12. CoicERichardGFHaberJE 2006 Saccharomyces cerevisiae donor preference during mating-type switching is dependent on chromosome architecture and organization. Genetics 173 1197 1206
13. CoicEMartinJRyuTTaySYKondevJ 2011 Dynamics of homology searching during gene conversion in Saccharomyces cerevisiae revealed by donor competition. Genetics 189 1225 1233
14. WeissKSimpsonRT 1997 Cell type-specific chromatin organization of the region that governs directionality of yeast mating type switching. Embo J 16 4352 4360
15. SunKCoicEZhouZDurrensPHaberJE 2002 Saccharomyces forkhead protein Fkh1 regulates donor preference during mating-type switching through the recombination enhancer. Genes Dev 16 2085 2096
16. CoicESunKWuCHaberJE 2006 Cell cycle-dependent regulation of Saccharomyces cerevisiae donor preference during mating-type switching by SBF (Swi4/Swi6) and Fkh1. Mol Cell Biol 26 5470 5480
17. ErcanSReeseJCWorkmanJLSimpsonRT 2005 Yeast recombination enhancer is stimulated by transcription activation. Mol Cell Biol 25 7976 7987
18. SzetoLFafaliosMKZhongHVershonAKBroachJR 1997 Alpha2p controls donor preference during mating type interconversion in yeast by inactivating a recombinational enhancer of chromosome III. Genes Dev 11 1899 1911
19. BystrickyKVan AttikumHMontielMDDionVGehlenL 2009 Regulation of nuclear positioning and dynamics of the silent mating type loci by the yeast Ku70/Ku80 complex. Mol Cell Biol 29 835 848
20. WeilerKSSzetoLBroachJR 1995 Mutations affecting donor preference during mating type interconversion in Saccharomyces cerevisiae. Genetics 139 1495 1510
21. TaddeiAHedigerFNeumannFRBauerCGasserSM 2004 Separation of silencing from perinuclear anchoring functions in yeast Ku80, Sir4 and Esc1 proteins. Embo J 23 1301 1312
22. DurocherDHenckelJFershtARJacksonSP 1999 The FHA domain is a modular phosphopeptide recognition motif. Mol Cell 4 387 394
23. DurocherDJacksonSP 2002 The FHA domain. FEBS Lett 513 58 66
24. LiJSmithGPWalkerJC 1999 Kinase interaction domain of kinase-associated protein phosphatase, a phosphoprotein-binding domain. Proc Natl Acad Sci U S A 96 7821 7826
25. LiaoHByeonIJTsaiMD 1999 Structure and function of a new phosphopeptide-binding domain containing the FHA2 of Rad53. J Mol Biol 294 1041 1049
26. LiJLeeGIVan DorenSRWalkerJC 2000 The FHA domain mediates phosphoprotein interactions. J Cell Sci 113 4143 4149
27. KimJAKruhlakMDotiwalaFNussenzweigAHaberJE 2007 Heterochromatin is refractory to gamma-H2AX modification in yeast and mammals. J Cell Biol 178 209 218
28. ShroffRArbel-EdenAPilchDIraGBonnerWM 2004 Distribution and dynamics of chromatin modification induced by a defined DNA double-strand break. Curr Biol 14 1703 1711
29. CheungWLTurnerFBKrishnamoorthyTWolnerBAhnSH 2005 Phosphorylation of histone H4 serine 1 during DNA damage requires casein kinase II in S. cerevisiae. Curr Biol 15 656 660
30. JainSSugawaraNLydeardJVazeMTanguy Le GacN 2009 A recombination execution checkpoint regulates the choice of homologous recombination pathway during DNA double-strand break repair. Genes Dev 23 291 303
31. LichtenMBortsRHHaberJE 1987 Meiotic gene conversion and crossing over between dispersed homologous sequences occurs frequently in Saccharomyces cerevisiae. Genetics 115 233 246
32. SugawaraNWangXHaberJE 2003 In vivo roles of Rad52, Rad54, and Rad55 proteins in Rad51-mediated recombination. Mol Cell 12 209 219
33. WolnerBvan KomenSSungPPetersonCL 2003 Recruitment of the recombinational repair machinery to a DNA double-strand break in yeast. Mol Cell 12 221 232
34. MooreJDYazganOAtaianYKrebsJE 2007 Diverse roles for histone H2A modifications in DNA damage response pathways in yeast. Genetics 176 15 25
35. HendzelMJWeiYManciniMAVan HooserARanalliT 1997 Mitosis-specific phosphorylation of histone H3 initiates primarily within pericentromeric heterochromatin during G2 and spreads in an ordered fashion coincident with mitotic chromosome condensation. Chromosoma 106 348 360
36. GotoHTomonoYAjiroKKosakoHFujitaM 1999 Identification of a novel phosphorylation site on histone H3 coupled with mitotic chromosome condensation. J Biol Chem 274 25543 25549
37. DaiJHigginsJM 2005 Haspin: a mitotic histone kinase required for metaphase chromosome alignment. Cell cycle 4 665 668
38. KaynePSKimUJHanMMullenJRYoshizakiF 1988 Extremely conserved histone H4 N terminus is dispensable for growth but essential for repressing the silent mating loci in yeast. Cell 55 27 39
39. TohGWSugawaraNDongJTothRLeeSE 2010 Mec1/Tel1-dependent phosphorylation of Slx4 stimulates Rad1-Rad10-dependent cleavage of non-homologous DNA tails. DNA repair 9 718 726
40. HannaDERethinaswamyAGloverCV 1995 Casein kinase II is required for cell cycle progression during G1 and G2/M in Saccharomyces cerevisiae. J Biol Chem 270 25905 25914
41. BressanDAVazquezJHaberJE 2004 Mating type-dependent constraints on the mobility of the left arm of yeast chromosome III. J Cell Biol 164 361 371
42. DekkerJRippeKDekkerMKlecknerN 2002 Capturing chromosome conformation. Science 295 1306 1311
43. MieleABystrickyKDekkerJ 2009 Yeast silent mating type loci form heterochromatic clusters through silencer protein-dependent long-range interactions. PLoS Genet 5 e1000478 doi:10.1371/journal.pgen.1000478
44. KnottSRPeaceJMOstrowAZGanYRexAE 2012 Forkhead transcription factors establish origin timing and long-range clustering in S. cerevisiae. Cell 148 99 111
45. BakkenistCJKastanMB 2003 DNA damage activates ATM through intermolecular autophosphorylation and dimer dissociation. Nature 421 499 506
46. BonillaCYMeloJAToczyskiDP 2008 Colocalization of sensors is sufficient to activate the DNA damage checkpoint in the absence of damage. Mol Cell 30 267 276
47. LeeJHPaullTT 2005 ATM activation by DNA double-strand breaks through the Mre11-Rad50-Nbs1 complex. Science 308 551 554
48. NakadaDMatsumotoKSugimotoK 2003 ATM-related Tel1 associates with double-strand breaks through an Xrs2-dependent mechanism. Genes Dev 17 1957 1962
49. CimprichKACortezD 2008 ATR: an essential regulator of genome integrity. Nat Rev Mol Cell Biol 9 616 627
50. ZouLElledgeSJ 2003 Sensing DNA damage through ATRIP recognition of RPA-ssDNA complexes. Science 300 1542 1548
51. DekkerJ 2008 Mapping in vivo chromatin interactions in yeast suggests an extended chromatin fiber with regional variation in compaction. J Biol Chem 283 34532 34540
52. EstojakJBrentRGolemisEA 1995 Correlation of two-hybrid affinity data with in vitro measurements. Mol Cell Biol 15 5820 5829
53. LeeSEMooreJKHolmesAUmezuKKolodnerRD 1998 Saccharomyces Ku70, mre11/rad50 and RPA proteins regulate adaptation to G2/M arrest after DNA damage. Cell 94 399 409
54. PaquesFLeungWYHaberJE 1998 Expansions and contractions in a tandem repeat induced by double-strand break repair. Mol Cell Biol 18 2045 2054
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