Isolation and genetic characterization of the Neurospora crassa REV1 and REV7 homologs: evidence for involvement in damage-induced mutagenesis.
In a previous paper, we reported that the Neurospora crassa upr-1 gene is a homolog of the yeast gene REV3, which encodes the catalytic subunit of DNA polymerase zeta (polzeta). Characterization of the upr-1 mutant indicated that the UPR1 protein plays a role in DNA repair and mutagenesis. To help understand the mechanisms of mutagenic DNA repair in the N. crassa more extensively, we identified N. crassa homologs of yeast REV1 and REV7 and obtained mutants ncrev1 or ncrev7, which had similar phenotypes to the upr-1 mutant. Mutant carrying ncrev7 was more sensitive to UV and 4NQO, and slightly sensitive to MMS than the wild-type. The sensitivity to UV and MMS of the ncrev1 mutant was moderately higher than that of the wild-type, but the sensitivity to 4NQO of the mutant was similar to that of the wild-type. In reversion assay using testers with base substitution or frameshift mutation at the ad-3A locus, each of ncrev1 and ncrev7 mutants showed lower induced-mutability than the wild-type. Expression of ncrev1 and ncrev7 was found to be UV-inducible like the case of upr-1. Genetic analyses showed that the ncrev7 was identical to mus-26, which belongs to the upr-1 epistasis group, and that the ncrev1 was a newly identified DNA repair gene and designated as mus-42. Interestingly, all three mutants have a normal CPD photolyase gene, however, they showed a partial photoreactivation defect (PPD) phenotype, not completely defective but inefficient in photoreactivation. These results suggest that N. crassa REV homolog genes function in DNA repair and UV mutagenesis through the bypass of (6-4) photoproducts.
Sakai
Wataru
W
Department of Regulation-Biology, Faculty of Science, Saitama University, City Saitama 338-8570, Japan.
Wada
Yuusaku
Y
Naoi
Yasuko
Y
Ishii
Chizu
C
Inoue
Hirokazu
H
D004247:DNA / Q000378:metabolism / Q000528:radiation effects; D004249:DNA Damage / Q000235:genetics; D004259:DNA-Directed DNA Polymerase / Q000235:genetics / Q000502:physiology; D004255:Deoxyribodipyrimidine Photo-Lyase / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics* / Q000302:isolation & purification* / Q000502:physiology; D009154:Mutation; D009492:Neurospora crassa / Q000235:genetics* / Q000502:physiology; D009713:Nucleotidyltransferases / Q000235:genetics / Q000502:physiology; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000502:physiology; D017385:Sequence Homology; D014466:Ultraviolet Rays; D000099270:Y-Family DNA Polymerases
D004247:DNA; D004259:DNA-Directed DNA Polymerase; D004255:Deoxyribodipyrimidine Photo-Lyase; D005656:Fungal Proteins; D009713:Nucleotidyltransferases; D029701:Saccharomyces cerevisiae Proteins; C092219:REV7 protein, S cerevisiae; C100951:REV1 protein, S cerevisiae; D000099270:Y-Family DNA Polymerases