Game J C, Kaufman P D
Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA.
Genetics. 1999 Feb;151(2):485-97. doi: 10.1093/genetics/151.2.485.
In vitro, the protein complex Chromatin Assembly Factor-I (CAF-I) from human or yeast cells deposits histones onto DNA templates after replication. In Saccharomyces cerevisiae, the CAC1, CAC2, and CAC3 genes encode the three CAF-I subunits. Deletion of any of the three CAC genes reduces telomeric gene silencing and confers an increase in sensitivity to killing by ultraviolet (UV) radiation. We used double and triple mutants involving cac1Delta and yeast repair gene mutations to show that deletion of the CAC1 gene increases the UV sensitivity of cells mutant in genes from each of the known DNA repair epistasis groups. For example, double mutants involving cac1Delta and excision repair gene deletions rad1Delta or rad14Delta showed increased UV sensitivity, as did double mutants involving cac1Delta and deletions of members of the RAD51 recombinational repair group. cac1Delta also increased the UV sensitivity of strains with defects in either the error-prone (rev3Delta) or error-free (pol30-46) branches of RAD6-mediated postreplicative DNA repair but did not substantially increase the sensitivity of strains carrying null mutations in the RAD6 or RAD18 genes. Deletion of CAC1 also increased the UV sensitivity and rate of UV-induced mutagenesis in rad5Delta mutants, as has been observed for mutants defective in error-free postreplicative repair. Together, these data suggest that CAF-I has a role in error-free postreplicative damage repair and may also have an auxiliary role in other repair mechanisms. Like the CAC genes, RAD6 is also required for gene silencing at telomeres. We find an increased loss of telomeric gene silencing in rad6Delta cac1Delta and rad18Delta cac1Delta double mutants, suggesting that CAF-I and multiple factors in the postreplicative repair pathway influence chromosome structure.
在体外,来自人或酵母细胞的染色质组装因子-I(CAF-I)蛋白复合物在复制后将组蛋白沉积到DNA模板上。在酿酒酵母中,CAC1、CAC2和CAC3基因编码CAF-I的三个亚基。三个CAC基因中任何一个的缺失都会降低端粒基因沉默,并导致对紫外线(UV)辐射杀伤的敏感性增加。我们使用涉及cac1Δ和酵母修复基因突变的双突变体和三突变体,以表明CAC1基因的缺失会增加来自每个已知DNA修复上位性组的基因突变体细胞的UV敏感性。例如,涉及cac1Δ和切除修复基因缺失rad1Δ或rad14Δ的双突变体显示出增加的UV敏感性,涉及cac1Δ和RAD51重组修复组成员缺失的双突变体也是如此。cac1Δ还增加了RAD6介导的复制后DNA修复的易错(rev3Δ)或无错(pol30-46)分支有缺陷的菌株的UV敏感性,但没有实质性增加携带RAD6或RAD18基因无效突变的菌株的敏感性。如在无错复制后修复缺陷的突变体中所观察到的,CAC1的缺失也增加了rad5Δ突变体中的UV敏感性和UV诱导的诱变率。总之,这些数据表明CAF-I在无错复制后损伤修复中起作用,并且可能在其他修复机制中也起辅助作用。与CAC基因一样,RAD6对于端粒处的基因沉默也是必需的。我们发现在rad6Δcac1Δ和rad18Δcac1Δ双突变体中端粒基因沉默的丧失增加,这表明CAF-I和复制后修复途径中的多种因子影响染色体结构。