CLONING AND CHARACTERIZATION OF RAD21 AN ESSENTIAL GENE OF SCHIZOSACCHAROMYCES-POMBE INVOLVED IN DNA DOUBLE-STRAND-BREAK REPAIR
CLONING AND CHARACTERIZATION OF RAD21 AN ESSENTIAL GENE OF SCHIZOSACCHAROMYCES-POMBE INVOLVED IN DNA DOUBLE-STRAND-BREAK REPAIR
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DOI：
10.1093/nar/20.24.6605
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发表时间：
1992-12-25
影响因子：
14.9
通讯作者：
SUBRAMANI, S
中科院分区：
文献类型：
--
作者：
BIRKENBIHL, RP;SUBRAMANI, S
通过脉冲场凝胶电泳对粟酒裂殖酵母染色体的分析表明，裂殖酵母具有非常有效的DNA双链断裂（dsb）修复系统，该系统在γ射线降解后适当地恢复了三条染色体。辐射敏感的突变体rad 21 -45是在这一修复途径的缺陷，但能够在G2期细胞周期阻滞DNA损伤。我们通过补充rad 21 -45突变体的辐射敏感性来克隆rad 21基因。质粒携带的基因完全重建了DNA双链断裂修复途径。通过杂交将rad 21基因定位于第1H染色体上。该转录本长2.5 kb，以中等水平表达。1884-bp的开放阅读框编码628个氨基酸，强酸性肽，计算分子量为67，854 D。rad 21基因与其他已知的核苷酸或肽序列没有显着的同源性。突变体不能进行有效的DNA修复是由单个碱基取代引起的，该碱基取代将突变体中的野生型异亮氨酸67变为苏氨酸。基因组rad 21基因的缺失表明，它是必不可少的有丝分裂的粟酒裂殖酵母的生长。
Analysis of the Schizosaccharomyces pombe chromosomes by pulsed field gel electrophoresis showed that the fission yeast has a very efficient DNA double-strand-break (dsb) repair system, which properly restores the three chromosomes after they are degraded by gamma-irradiation. The radiation-sensitive mutant rad21-45 is deficient in this repair pathway but is capable of cell-cycle arrest in G2 following DNA damage. We cloned the rad21 gene by complementing the radiation sensitivity of the rad21-45 mutant. The plasmid-borne gene completely reestablished the DNA dsb repair pathway. The rad21 gene was localized to chromosome IH by hybridization. The transcript is 2.5 kb long and expressed at a moderate level. The 1884-bp open reading frame encodes a 628 amino acid, very acidic peptide with a calculated molecular mass of 67,854 D. The rad21 gene shows no significant homology to other known nucleotide or peptide sequences. The inability of the mutant to perform efficient DNA repair is caused by a single base substitution, which changes wild-type isoleucine67 into threonine in the mutant. Deletion of the genomic rad21 gene showed that it is essential for mitotic growth of S.pombe.