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酿酒酵母一种新的DNA损伤诱导基因DIN7的特性分析,该基因是RAD2和RAD27 DNA修复基因的结构同源物。

Characterization of a novel DNA damage-inducible gene of Saccharomyces cerevisiae, DIN7, which is a structural homolog of the RAD2 and RAD27 DNA repair genes.

作者信息

Mieczkowski P A, Fikus M U, Ciesla Z

机构信息

Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw.

出版信息

Mol Gen Genet. 1997 Feb 27;253(6):655-65. doi: 10.1007/s004380050369.

Abstract

A number of DNA damage-inducible genes (DIN) have been identified in Saccharomyces cerevisiae. In the present study we describe isolation of a novel gene, Din7, the expression of which is induced by exposure of cells to UV light, MMS (methyl methane-sulfonate) or HU (hydoxyurea). The DNA sequence of DIN7 was determined. By comparison of the predicted Din7 amino acid sequence with those in databases we found that it belongs to a family of proteins which includes S. cerevisiae Rad2 and its Schizosaccharomyces pombe and human homologs Rad13 and XPGC; S. cerevisiae Rad27 and its S. pombe homolog Rad2, and S. pombe Exo I. All these proteins are endowed with DNA nuclease activity and are known to play an important function in DNA repair. The strongest homology to Din7 was found with the Dhs1 protein of S. cerevisiae, the function of which is essentially unknown. The expression of the DIN7 gene was studied in detail using a DIN7-lacZ fusion integrated into a chromosome. We show that the expression level of DIN7 rises during meiosis at a time nearly coincident with commitment to recombination. No inducibility of DIN7 was found after treatment with DNA-damaging agents of cells bearing the rad53-21 mutation. Surprisingly, a high basal level of DIN7 expression was found in strains in which the DUN1 gene was inactivated by transposon insertion. We suggest that a form of Dun1 may be a negative regulator of the DIN7 gene expression.

摘要

在酿酒酵母中已鉴定出许多DNA损伤诱导基因(DIN)。在本研究中,我们描述了一个新基因Din7的分离,该基因的表达可通过将细胞暴露于紫外线、甲磺酸甲酯(MMS)或羟基脲(HU)来诱导。测定了DIN7的DNA序列。通过将预测的Din7氨基酸序列与数据库中的序列进行比较,我们发现它属于一个蛋白质家族,该家族包括酿酒酵母Rad2及其粟酒裂殖酵母和人类同源物Rad13和XPGC;酿酒酵母Rad27及其粟酒裂殖酵母同源物Rad2,以及粟酒裂殖酵母Exo I。所有这些蛋白质都具有DNA核酸酶活性,并且已知在DNA修复中起重要作用。与Din7同源性最强的是酿酒酵母的Dhs1蛋白,其功能基本未知。使用整合到染色体中的DIN7-lacZ融合体详细研究了DIN7基因的表达。我们表明,DIN7的表达水平在减数分裂期间升高,时间几乎与重组开始一致。在用rad53-21突变的细胞进行DNA损伤剂处理后,未发现DIN7的诱导性。令人惊讶的是,在通过转座子插入使DUN1基因失活的菌株中发现了高水平的DIN7基础表达。我们认为Dun1的一种形式可能是DIN7基因表达的负调节因子。

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