Isolation and genetic characterisation of the Drosophila homologue of (SCE)REV3, encoding the catalytic subunit of DNA polymerase zeta.
In Drosophila, about 30 mutants are known that show hypersensitivity to the methylating agent methyl methane sulfonate (MMS). 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.
Eeken
J C
JC
Department of Radiation Genetics and Chemical Mutagenesis, MGC, Leiden University Medical Center, P.O. Box 9503, 2300 RF, Leiden, The Netherlands. j.c.j.eeken@lumc.nl
Romeijn
R J
RJ
de Jong
A W
AW
Pastink
A
A
Lohman
P H
PH
D000595:Amino Acid Sequence; D000818:Animals; D020134:Catalytic Domain; D003001:Cloning, Molecular; D004258:DNA Polymerase III / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D004259:DNA-Directed DNA Polymerase / Q000235:genetics*; D004331:Drosophila melanogaster / Q000201:enzymology / Q000235:genetics*; D017344:Genes, Insect; D008741:Methyl Methanesulfonate / Q000494:pharmacology; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009153:Mutagens / Q000494:pharmacology; D009154:Mutation; D020161:Physical Chromosome Mapping; D021122:Protein Subunits; D011836:Radiation Tolerance / Q000235:genetics; D011995:Recombination, Genetic; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D009153:Mutagens; D021122:Protein Subunits; D008741:Methyl Methanesulfonate; C425929:DNA polymerase zeta; D004258:DNA Polymerase III; D004259:DNA-Directed DNA Polymerase