Disruption of mouse <i>SNM1</i> causes increased sensitivity to the DNA interstrand cross-linking agent mitomycin C

作     者：Dronkert, MLG de Wit, J Boeve, M Vasconcelos, ML van Steeg, H Tan, TLR Hoeijmakers, JHJ Kanaar, R 

作者机构：Erasmus Univ Dept Cell Biol & Genet Ctr Biomed Genet NL-3000 DR Rotterdam Netherlands Daniel den Hoed Canc Ctr Dept Radiat Oncol NL-3000 DR Rotterdam Netherlands Natl Inst Publ Hlth & Environm Hlth Effects Res Lab NL-3720 BA Bilthoven Netherlands 

出 版 物：《MOLECULAR AND CELLULAR BIOLOGY》 (分子生物学与细胞生物学)

年 卷 期：2000年第20卷第13期

页      面：4553-4561页

核心收录：

学科分类：0710[理学-生物学] 071010[理学-生物化学与分子生物学] 07[理学] 

主　　题：氨基酸序列 交联试剂/药理学 DNA修复/药物作用 DNA结合蛋白质类/药物作用 DNA结合蛋白质类/遗传学 DNA结合蛋白质类/代谢 药物过敏/遗传学 内切脱氧核糖核酸酶类 成纤维细胞 小鼠 近交C57BL 小鼠 基因敲除 丝裂霉素/药理学 分子序列数据 核蛋白质类/药物作用 核蛋白质类/遗传学 核蛋白质类/代谢 酿酒酵母蛋白质类 序列同源性 氨基酸 干细胞/生理学 亚细胞部分 动物 女(雌)性 人类 男(雄)性 小鼠 

摘      要：DNA interstrand cross-links (ICLs) represent lethal DNA damage, because they block transcription, replication, and segregation of DNA. Because of their genotoxicity, agents inducing ICLs are often used in antitumor therapy. The repair of ICLs is complex and involves proteins belonging to nucleotide excision, recombination, and translesion DNA repair pathways in Escherichia coli, Saccharomyces cerevisiae, and mammals. We cloned and analyzed mammalian homologs of the S. cerevisiae gene SNM1 (PSO2), which is specifically involved in ICL repair. Human Snm1, a nuclear protein, was ubiquitously expressed at a very low level. We generated mouse SNM1(-/-) embryonic stem cells and showed that these cells were sensitive to mitomycin C. In contrast to S. cerevisiae snm1 mutants, they were not significantly sensitive to other ICL agents, probably due to redundancy in mammalian ICL repair and the existence of other SNM1 homologs. The sensitivity to mitomycin C was complemented by transfection of the human SNM1 cDNA and by targeting of a genomic cDNA-murine SNM1 fusion construct to the disrupted locus. We also generated mice deficient for murine SNM1. They were viable and fertile and showed no major abnormalities. However, they were sensitive to mitomycin C. The ICL sensitivity of the mammalian SNM1 mutant suggests that SNM1 function and, by implication, ICL repair are at least partially conserved between S. cerevisiae and mammals.