Abstract
The rev6-1 allele was isolated in a screen for mutants deficient for UV-induced reversion of the frameshift mutation his4-38. Preliminary testing showed that the rev6-1 mutant was substantially deficient for UV-induced reversion of arg4-17 and ilv1-92 and markedly UV sensitive. Unlike other REV genes, which encode DNA polymerases and an associated subunit, REV6 has been found to be identical to POL30, which encodes proliferating cell nuclear antigen (PCNA), the subunit of the homotrimeric sliding clamp, in which the rev6-1 mutation produces a G178S substitution. This substitution appears to abolish all DNA damage-tolerance activities normally carried out by the RAD6/RAD18 pathway, including translesion replication by DNA polymerase zeta/Rev1 and DNA polymerase eta, and the error-free, recombination-dependent component of this pathway, but has little effect on the growth rate, suggesting that G178S may prevent ubiquitination of lysine 164 in PCNA. We also find that rev6-1 mutation can be fully complemented by a centromere-containing, low copy-number plasmid carrying POL30, despite the presumed occurrence in the mutant of sliding clamp assemblies that contain between one and three G178S PCNA monomers as well as the fully wild-type species.
MeSH Terms
Alleles
Amino Acid Substitution
DNA Damage/genetics,radiation effects
DNA-Directed DNA Polymerase/genetics,metabolism
Genetic Complementation Test
Plasmids/genetics,metabolism
Point Mutation
Proliferating Cell Nuclear Antigen/genetics,metabolism
Recombination, Genetic
Saccharomyces cerevisiae/genetics,growth & development,radiation effects
Saccharomyces cerevisiae Proteins/genetics,metabolism
Ultraviolet Rays
Chemicals
Proliferating Cell Nuclear Antigen
Saccharomyces cerevisiae Proteins
DNA-Directed DNA Polymerase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Zhang Hengshan
Department of Biochemistry and Biophysics, University of Rochester School of Medicine and Dentistry, Rochester, New York 14642, USA.
Gibbs Peter E M
Lawrence Christopher W
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