The Schizosaccharomyces pombe rad60 gene is essential for repairing double-strand DNA breaks spontaneously occurring during replication and induced by DNA-damaging agents
The Schizosaccharomyces pombe rad60 gene is essential for repairing double-strand DNA breaks spontaneously occurring during replication and induced by DNA-damaging agents
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DOI：
10.1128/mcb.22.10.3537-3548.2002
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发表时间：
2002-05-01
影响因子：
5.3
通讯作者：
Shinagawa, H
中科院分区：
文献类型：
--
作者：
Morishita, T;Tsutsui, Y;Shinagawa, H
为了确定新的基因参与DNA双链断裂（DSB）修复，我们以前分离裂殖酵母粟酒裂殖酵母突变体，这是超敏感的甲基甲磺酸盐（MMS）和合成致死rad 2。本研究的特点，这些突变体之一，rad 60 -1。克隆了补充该突变体的MMS敏感性的基因，并命名为rad 60。RAD 60编码具有406个氨基酸的蛋白质，其具有在遍在蛋白家族蛋白质中发现的保守的遍在蛋白-2基序。rad 60 -1对紫外线和γ射线超敏，对rhp 51上位，并且在γ-辐射引起的DSB的修复中有缺陷。rad 60 -1突变体对生长也是温度敏感的。在限制性温度（37 ℃）下，rad 60 -1细胞生长几次分裂，然后用2C DNA内容物停滞;停滞的细胞积累DSB，并具有扩散的且通常形状异常的核染色体结构域。rad 60 -1突变体与rad 18-X是合成致死的，并且来自多拷贝质粒的野生型rad 60的表达部分地抑制了rad 18-X细胞的MMS敏感性。rad 18编码染色体结构维持（SMC）家族（A. R. Lehmann，M. Walicka，D.格里菲思，J.M.默里角，加-地Z. Watts，S. McCready，和A. M.卡尔，摩尔Cell. 15：7067-7080，1995）。这些结果提示S.粟酒酵母Rad 60需要通过姐妹染色单体之间的重组来修复在复制过程中积累的DSB。Rad 60可能与SMC蛋白Rad 18一起执行此功能。
To identify novel genes involved in DNA double-strand break (DSB) repair, we previously isolated Schizosaccharomyces pombe mutants which are hypersensitive to methyl methanesulfonate (MMS) and synthetic lethals with rad2. This study characterizes one of these mutants, rad60-1. The gene that complements the MMS sensitivity of this mutant was cloned and designated rad60. rad60 encodes a protein with 406 amino acids which has the conserved ubiquitin-2 motif found in ubiquitin family proteins. rad60-1 is hypersensitive to UV and gamma rays, epistatic to rhp51, and defective in the repair of DSBs caused by gamma-irradiation. The rad60-1 mutant is also temperature sensitive for growth. At the restrictive temperature (37 C), rad60-1 cells grow for several divisions and then arrest with 2C DNA content; the arrested cells accumulate DSBs and have a diffuse and often aberrantly shaped nuclear chromosomal domain. The rad60-1 mutant is a synthetic lethal with rad18-X, and expression of wild-type rad60 from a multicopy plasmid partially suppresses the MMS sensitivity of rad18-X cells. rad18 encodes a conserved protein of the structural maintenance of chromosomes (SMC) family (A. R. Lehmann, M. Walicka, D. J. Griffiths, J. M. Murray, F. Z. Watts, S. McCready, and A. M. Carr, Mol. Cell. Biol. 15:7067-7080, 1995). These results suggest that S. pombe Rad60 is required to repair DSBs, which accumulate during replication, by recombination between sister chromatids. Rad60 may perform this function in concert with the SMC protein Rad18.