Friedberg E C, Fleer R, Naumovski L, Nicolet C M, Robinson G W, Weiss W A, Yang E
Basic Life Sci. 1986;39:231-42. doi: 10.1007/978-1-4684-5182-5_20.
The genetics of nucleotide excision repair in the yeast Saccharomyces cerevisiae is complex, apparently requiring at least 10 genes. We have isolated 5 of these genes (designated RAD1, RAD2, RAD3, RAD4, and RAD10) by molecular cloning and plan to overexpress them in order to generate proteins for biochemical study. We have sequenced four of these five genes and have noted regions of homology with other proteins in the predicted amino acid sequence of some of them. In particular, there is striking homology between Rad3 protein and a number of prokaryotic and eukaryotic proteins that bind nucleotides and hydrolyze ATP or GTP. Mutations in this region of the RAD3 gene render cells defective in the nucleotide excision repair function. In addition to its role in nucleotide excision repair, the RAD3 gene is essential for the viability of haploid cells in the absence of DNA damage. The nature of the essential function is unknown. The RAD1 and RAD3 genes are not inducible by DNA damaging agents. However, exposure of cells to UV radiation, 4-nitroquinoline 1-oxide, or gamma radiation results in 4- to 6-fold enhanced expression of the RAD2 gene.
酿酒酵母中核苷酸切除修复的遗传学机制很复杂,显然至少需要10个基因。我们通过分子克隆分离出了其中5个基因(命名为RAD1、RAD2、RAD3、RAD4和RAD10),并计划对它们进行过表达,以产生用于生化研究的蛋白质。我们已经对这5个基因中的4个进行了测序,并在其中一些基因的预测氨基酸序列中发现了与其他蛋白质的同源区域。特别是,Rad3蛋白与许多结合核苷酸并水解ATP或GTP的原核和真核蛋白质之间存在显著的同源性。RAD3基因这一区域的突变会使细胞在核苷酸切除修复功能上出现缺陷。除了在核苷酸切除修复中的作用外,RAD3基因对于单倍体细胞在无DNA损伤情况下的生存能力也是必需的。其必需功能的本质尚不清楚。RAD1和RAD3基因不受DNA损伤剂的诱导。然而,将细胞暴露于紫外线、4-硝基喹啉1-氧化物或γ辐射下会导致RAD2基因的表达增强4至6倍。