Isolation and genetic characterisation of the Drosophila homologue of (SCE)REV3, encoding the catalytic subunit of DNA polymerase ζ

被引:20
作者
Eeken, JCJ
Romeijn, RJ
de Jong, AWM
Pastink, A
Lohman, PHM
机构
[1] Leiden Univ, Med Ctr, MGC, Dept Radiat Genet & Chem Mutagenesis, NL-2300 RF Leiden, Netherlands
[2] JA Cohen Inst, Interuniv Res Inst Radiopathol & Radiat Protect, Leiden, Netherlands
来源
MUTATION RESEARCH-DNA REPAIR | 2001年 / 485卷 / 03期
关键词
translesion synthesis; Drosophila; mus205; REV3;
D O I
10.1016/S0921-8777(01)00062-3
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
In Drosophila, about 30 mutants are known that show hypersensitivity to the methylating agent methyl methane sulfonate CMMS). Addition of this agent to the medium results in an increased larval mortality of the mutants. Using a P-insertion mutagenesis screen, three MMS-sensitive mutants on chromosome II were isolated. One of these is allelic to the known EMS-induced mus205 (mutagen sensitive) mutant. In the newly isolated mutant, a P-element is detected in region 43E by in situ hybridisation. The localisation of mus205 to this region was confirmed by deficiency mapping. The gene was cloned and shows strong homology to the Saccharomyces cerevisiae REV3 gene. The REV3 gene encodes the catalytic subunit of DNA polymerase zeta, involved in translesion synthesis. The P-element is inserted in the first exon of the mus205 gene resulting in an aberrant mRNA, encoding a putative truncated protein containing only the first 13 of the 2130 aa native Drosophila protein. The mus205 mutant is hypersensitive to alkylating agents and UV, but not to ionising radiation. In contrast to reported data, in germ cells, the mutant has no effect on mutability by X-rays, NQO and alkylating agents. In somatic cells, the mutant shows no effect on MMS-induced mutations and recombinations. This phenotype of the Drosophila mus205 mutant is strikingly different from the phenotype of the yeast rev3 mutant, which is hypomutable after UV, X-rays, NQO and alkylating agents. (C) 2001 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:237 / 253
页数:17
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