RRC ID 15692
著者 Tanae K, Horiuchi T, Matsuo Y, Katayama S, Kawamukai M.
タイトル Histone chaperone Asf1 plays an essential role in maintaining genomic stability in fission yeast.
ジャーナル PLoS One
Abstract The histone H3-H4 chaperone Asf1 is involved in chromatin assembly (or disassembly), histone exchange, regulation of transcription, and chromatin silencing in several organisms. To investigate the essential functions of Asf1 in Schizosaccharomyces pombe, asf1-ts mutants were constructed by random mutagenesis using PCR. One mutant (asf1-33(ts)) was mated with mutants in 77 different kinase genes to identify synthetic lethal combinations. The asf1-33 mutant required the DNA damage checkpoint factors Chk1 and Rad3 for its survival at the restrictive temperature. Chk1, but not Cds1, was phosphorylated in the asf1-33 mutant at the restrictive temperature, indicating that the DNA damage checkpoint was activated in the asf1-33 mutant. DNA damage occured in the asf1-33 mutant, with degradation of the chromosomal DNA observed through pulse-field gel electrophoresis and the formation of Rad22 foci. Sensitivity to micrococcal nuclease in the asf1-33 mutant was increased compared to the asf1(+) strain at the restrictive temperature, suggesting that asf1 mutations also caused a defect in overall chromatin structure. The Asf1-33 mutant protein was mislocalized and incapable of binding histones. Furthermore, histone H3 levels at the centromeric outer repeat region were decreased in the asf1-33 mutant and heterochromatin structure was impaired. Finally, sim3, which encodes a CenH3 histone chaperone, was identified as a strong suppressor of the asf1-33 mutant. Taken together, these results clearly indicate that Asf1 plays an essential role in maintaining genomic stability in S. pombe.
巻・号 7(1)
ページ e30472
公開日 2012-1-1
DOI 10.1371/journal.pone.0030472
PII PONE-D-11-16975
PMID 22291963
PMC PMC3266922
MeSH Adaptation, Physiological / genetics Cell Cycle Checkpoints / genetics Cell Cycle Proteins / genetics Cell Cycle Proteins / physiology Cell Shape / genetics Cell Survival / genetics Checkpoint Kinase 1 Checkpoint Kinase 2 Genomic Instability / genetics* Histone Chaperones / genetics Histone Chaperones / metabolism Histone Chaperones / physiology Histones / metabolism Models, Biological Molecular Chaperones / genetics Molecular Chaperones / metabolism Molecular Chaperones / physiology* Organisms, Genetically Modified Protein Binding / genetics Protein Kinases / genetics Protein Kinases / physiology Schizosaccharomyces / cytology Schizosaccharomyces / genetics* Schizosaccharomyces / physiology Schizosaccharomyces pombe Proteins / genetics Schizosaccharomyces pombe Proteins / metabolism Schizosaccharomyces pombe Proteins / physiology* Temperature
IF 2.74
引用数 9
WOS 分野 BIOCHEMISTRY & MOLECULAR BIOLOGY
リソース情報
酵母 FY14069