Doe C L, Murray J M, Shayeghi M, Hoskins M, Lehmann A R, Carr A M, Watts F Z
School of Biological Sciences, University of Sussex, Falmer, Brighton, UK.
Nucleic Acids Res. 1993 Dec 25;21(25):5964-71. doi: 10.1093/nar/21.25.5964.
The Schizosaccharomyces pombe rad8 mutant is sensitive to both UV and gamma irradiation. We have cloned the rad8 gene by complementation of the UV sensitivity of a rad8.190 mutant strain. The gene comprises an open reading frame of 3.4 kb which does not contain any introns and is capable of encoding a 1133 amino acid protein of 129 kDa. Deletion of the gene indicates that it is not essential for cell viability. Recognisable motifs are present for a nuclear localisation signal, a RING finger and helicase domains. The predicted protein is a member of the SNF2 subfamily of proteins and shows particular homology to the Saccharomyces cerevisiae RAD5 protein. Double mutant analysis demonstrated that the rad8 mutant is not epistatic to mutants in the excision repair pathway (rad13) or checkpoint pathway (rad9). Analysis of radiation sensitivity though the cell cycle indicates that, unlike most other rad mutants, rad8 is most sensitive to irradiation during the G1/S period.
粟酒裂殖酵母rad8突变体对紫外线和γ射线照射均敏感。我们通过互补rad8.190突变菌株的紫外线敏感性克隆了rad8基因。该基因包含一个3.4kb的开放阅读框，不含任何内含子，能够编码一个129kDa的1133个氨基酸的蛋白质。该基因的缺失表明它对细胞活力并非必需。存在可识别的核定位信号、RING指结构域和解旋酶结构域基序。预测的蛋白质是SNF2蛋白质亚家族的成员，与酿酒酵母RAD5蛋白具有特别的同源性。双突变分析表明，rad8突变体在切除修复途径（rad13）或检查点途径（rad9）中对突变体不具有上位性。通过细胞周期分析辐射敏感性表明，与大多数其他rad突变体不同，rad8在G1/S期对辐射最敏感。
