Multiple roles of Rev3, the catalytic subunit of polzeta in maintaining genome stability in vertebrates.
Translesion DNA synthesis (TLS) and homologous DNA recombination (HR) are two major postreplicational repair (PRR) pathways. The REV3 gene of Saccharomyces cerevisiae encodes the catalytic subunit of DNA polymerase zeta, which is involved in mutagenic TLS. To investigate the role of REV3 in vertebrates, we disruped the gene in chicken DT40 cells. REV3(-/-) cells are sensitive to various DNA-damaging agents, including UV, methyl methanesulphonate (MMS), cisplatin and ionizing radiation (IR), consistent with its role in TLS. Interestingly, REV3(-/-) cells showed reduced gene targeting efficiencies and significant increase in the level of chromosomal breaks in the subsequent M phase after IR in the G(2) phase, suggesting the involvement of Rev3 in HR-mediated double-strand break repair. REV3(-/-) cells showed significant increase in sister chromatid exchange events and chromosomal breaks even in the absence of exogenous genotoxic stress. Furthermore, double mutants of REV3 and RAD54, genes involved in HR, are synthetic lethal. In conclusion, Rev3 plays critical roles in PRR, which accounts for survival on naturally occurring endogenous as well as induced damages during replication.
Sonoda
Eiichiro
E
CREST Research Project, Japan Science and Technology, Radiation Genetics, Graduate School of Medicine, Kyoto University, Yoshida Konoe, Sakyo-ku, Kyoto 606-8501, Japan.
Okada
Takashi
T
Zhao
Guang Yu
GY
Tateishi
Satoshi
S
Araki
Kasumi
K
Yamaizumi
Masaru
M
Yagi
Takashi
T
Verkaik
Nicole S
NS
van Gent
Dik C
DC
Takata
Minoru
M
Takeda
Shunichi
S
D000818:Animals; D020134:Catalytic Domain; D002460:Cell Line; D002645:Chickens; D002869:Chromosome Aberrations; D004247:DNA / Q000235:genetics* / Q000378:metabolism / Q000528:radiation effects; D004249:DNA Damage; D004265:DNA Helicases; D004260:DNA Repair; D045643:DNA Repair Enzymes; D004261:DNA Replication / Q000502:physiology*; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016678:Genome; D009154:Mutation; D011839:Radiation, Ionizing; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012854:Sister Chromatid Exchange
D029701:Saccharomyces cerevisiae Proteins; D004247:DNA; C425929:DNA polymerase zeta; D004259:DNA-Directed DNA Polymerase; C092602:REV3 protein, S cerevisiae; C075468:RAD54 protein, S cerevisiae; D004265:DNA Helicases; D045643:DNA Repair Enzymes