Carr A M, Sheldrick K S, Murray J M, al-Harithy R, Watts F Z, Lehmann A R
MRC Cell Mutation Unit, Sussex University, Falmer, Brighton, UK.
Nucleic Acids Res. 1993 Mar 25;21(6):1345-9. doi: 10.1093/nar/21.6.1345.
Cells mutated at the rad13 locus in the fission yeast, Schizosaccharomyces pombe are deficient in excision-repair of UV damage. We have cloned the S.pombe rad13 gene by its ability to complement the UV sensitivity of a rad13 mutant. The gene is not essential for cell proliferation. Sequence analysis of the cloned gene revealed an open reading-frame of 1113 amino acids with structural homology to the RAD2 gene of the distantly related Saccharomyces cerevisiae. The sequence similarity is confined to three domains, two close to the N-terminus of the encoded protein, the third being close to the C-terminus. The central region of about 500 amino acids shows little similarity between the two organisms. The first and third domains are also found in a related yet distinct pair of homologous S.pombe/S.cerevisiae DNA repair genes (rad2/YKL510), which have only a very short region between these two conserved domains. Using the polymerase chain reaction with degenerate primers, we have isolated fragments from a gene homologous to rad13/RAD2 from Aspergillus nidulans. These findings define new functional domains involved in excision-repair, as well as identifying a conserved family of genes related to RAD2.
裂殖酵母粟酒裂殖酵母中rad13基因座发生突变的细胞在紫外线损伤的切除修复方面存在缺陷。我们通过其互补rad13突变体紫外线敏感性的能力克隆了粟酒裂殖酵母rad13基因。该基因对细胞增殖不是必需的。对克隆基因的序列分析揭示了一个由1113个氨基酸组成的开放阅读框，与远缘的酿酒酵母的RAD2基因具有结构同源性。序列相似性局限于三个结构域，两个靠近编码蛋白的N端，第三个靠近C端。约500个氨基酸的中央区域在这两种生物之间几乎没有相似性。第一和第三结构域也存在于一对相关但不同的粟酒裂殖酵母/酿酒酵母同源DNA修复基因（rad2/YKL510）中，这两个保守结构域之间只有非常短的区域。使用简并引物进行聚合酶链反应，我们从构巢曲霉中分离出了与rad13/RAD2同源的基因片段。这些发现定义了参与切除修复的新功能结构域，并鉴定了与RAD2相关的保守基因家族。
