Deletion of the chromatin remodeling gene SPT10 sensitizes yeast cells to a subclass of DNA-damaging agents.
The Saccharomyces cerevisiae SPT10 protein possesses a DNA-binding domain that is fused to a putative histone acetyltransferase domain. It binds specifically to upstream-activating sequence elements in the core histone promoters and plays a direct role in histone gene regulation. SPT10 is also required for cell-cycle-specific K56 acetylation at histone genes, allowing the recruitment of the nucleosome remodeling factor Snf5 and subsequent regulation of gene transcription. We reisolated the SPT10 gene in a functional genome-wide screen designed to identify haploid yeast mutants that are hypersensitive to the antitumor drug bleomycin, which acts by damaging DNA. In addition to bleomycin, we show that spt10Delta mutants are also hypersensitive to a limited set of genotoxic agents that create DNA strand breaks, but not to 254-nm ultraviolet light or 4-nitroquinoline-1-oxide, which generate helix distortion. The hypersensitivities of the spt10Delta mutant to the genotoxic agents are rescued by a single copy plasmid carrying the SPT10 gene. We further showed that spt10Delta mutants displayed a modest twofold increase spontaneous mutant frequency, as compared to the parent. Following exposure to bleomycin, these mutants accumulate unrepaired lesions, e.g., DNA strand breaks with blocked 3'-ends in the chromosomal DNA. This defect is not due to the altered expression level or the enzymatic activities of a key DNA repair enzyme, APN1, which is known to repair DNA strand breaks with blocked ends. We propose that SPT10 mediates repair of a subset of DNA lesions by acetylating histones to promote recruitment of DNA repair enzymes.
Tounekti
Kaouther
K
Laboratoire de Biochimie et de Biotechnologie, Faculte des Sciences de Tunis, Université Tunis El-Manar, Tunis, Tunisia.
Aouida
Mustapha
M
Leduc
Anick
A
Poschmann
Jeremie
J
Yang
Xiaoming
X
Belhadj
Omrane
O
Ramotar
Dindial
D
D015112:4-Nitroquinoline-1-oxide / Q000633:toxicity; D001761:Bleomycin / Q000633:toxicity; D002843:Chromatin / Q000235:genetics; D004249:DNA Damage; D004260:DNA Repair; D005720:Gamma Rays / Q000009:adverse effects; D051548:Histone Acetyltransferases; D006861:Hydrogen Peroxide / Q000633:toxicity; D008741:Methyl Methanesulfonate / Q000633:toxicity; D009153:Mutagens / Q000633:toxicity*; D010692:Phleomycins / Q000633:toxicity; D015363:Quinolones / Q000633:toxicity; D051135:Rad51 Recombinase / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000187:drug effects* / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics*; D014466:Ultraviolet Rays / Q000009:adverse effects
C120195:4-nitroquinolone-1-oxide; D002843:Chromatin; D009153:Mutagens; D010692:Phleomycins; D015363:Quinolones; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D001761:Bleomycin; D015112:4-Nitroquinoline-1-oxide; D008741:Methyl Methanesulfonate; D006861:Hydrogen Peroxide; D051548:Histone Acetyltransferases; C086444:SPT10 protein, S cerevisiae; C495151:RAD51 protein, S cerevisiae; D051135:Rad51 Recombinase