Zhang Hengshan, Gibbs Peter E M, Lawrence Christopher W
Department of Biochemistry and Biophysics, University of Rochester School of Medicine and Dentistry, Rochester, New York 14642, USA.
Genetics. 2006 Aug;173(4):1983-9. doi: 10.1534/genetics.106.058545. Epub 2006 Jun 18.
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.
rev6-1等位基因是在筛选对紫外线诱导的移码突变his4-38回复突变缺陷的突变体时分离得到的。初步测试表明,rev6-1突变体对紫外线诱导的arg4-17和ilv1-92回复突变严重缺陷,且对紫外线明显敏感。与其他编码DNA聚合酶和相关亚基的REV基因不同,REV6已被发现与POL30相同,POL30编码增殖细胞核抗原(PCNA),即同三聚体滑动夹钳的亚基,其中rev6-1突变产生G178S替代。这种替代似乎消除了RAD6/RAD18途径通常进行的所有DNA损伤耐受活性,包括DNA聚合酶zeta/Rev1和DNA聚合酶eta的跨损伤复制,以及该途径的无差错、依赖重组的成分,但对生长速率影响很小,这表明G178S可能阻止PCNA中赖氨酸164的泛素化。我们还发现,尽管推测突变体中存在含有一到三个G178S PCNA单体以及完全野生型的滑动夹钳组装体,但携带POL30的含着丝粒的低拷贝数质粒可以完全互补rev6-1突变。