The Saccharomyces cerevisiae rev6-1 mutation, which inhibits both the lesion bypass and the recombination mode of DNA damage tolerance, is an allele of POL30, encoding proliferating cell nuclear antigen - PubMed (original) (raw)

The Saccharomyces cerevisiae rev6-1 mutation, which inhibits both the lesion bypass and the recombination mode of DNA damage tolerance, is an allele of POL30, encoding proliferating cell nuclear antigen

Hengshan Zhang et al. Genetics. 2006 Aug.

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.

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Figures

Figure 1.—

Frequencies of UV-induced reversion of arg4-17 (A, C, and E) and percentage of survival following UV irradiation (B, D, and F) in REV6, rev6-1, _rev1_Δ, _rev3_Δ, _rad30_Δ, and _pol32_Δ strains and in _rev6-1 rev1_Δ, _rev6-1 rev3_Δ, _rev6-1 rad30_Δ, and _rev6-1 pol32_Δ double mutants. A–F: ▪, REV6+; *, rev6-1. Additionally, in A and B: ○, _rad30_Δ; ▿, _pol32_Δ; •, _rev1_Δ; ▾, _rev3_Δ. In C and D: ○, _rev6-1 rev1_Δ; ▿, _rev6-1 pol32_Δ ; •, _rev6-1 rad30_Δ; ▾, _rev6-1 rev3_Δ. In E and F: ○, rev6-1 + YCplac33 POL30; ▾, rev6-1 + YCplac33. Data are the average of two or three replicate experiments.

Figure 2.—

Growth rate monitored by turbidity (arbitrary absorbance units) and viable cell titer in REV6+ and rev6-1 strains transformed with either YCplac33 POL30 or YCplac33. ▪, REV6; *, rev6-1; ○, rev6-1 with YCplac33 POL30; ▾, rev6-1 with YCplac33. Data are the average of three experiments.

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The Saccharomyces cerevisiae rev6-1 mutation, which inhibits both the lesion bypass and the recombination mode of DNA damage tolerance, is an allele of POL30, encoding proliferating cell nuclear antigen - PubMed (original) (raw)