title: Both high-fidelity replicative and low-fidelity Y-family polymerases are involved in DNA rereplication.
abstract: DNA rereplication is a major form of aberrant replication that causes genomic instabilities, such as gene amplification. However, little is known about which DNA polymerases are involved in the process. Here, we report that low-fidelity Y-family polymerases (Y-Pols), Pol η, Pol ι, Pol κ, and REV1, significantly contribute to DNA synthesis during rereplication, while the replicative polymerases, Pol δ and Pol ε, play an important role in rereplication, as expected. When rereplication was induced by depletion of geminin, these polymerases were recruited to rereplication sites in human cell lines. This finding was supported by RNA interference (RNAi)-mediated knockdown of the polymerases, which suppressed rereplication induced by geminin depletion. Interestingly, epistatic analysis indicated that Y-Pols collaborate in a common pathway, independently of replicative polymerases. We also provide evidence for a catalytic role for Pol η and the involvement of Pol η and Pol κ in cyclin E-induced rereplication. Collectively, our findings indicate that, unlike normal S-phase replication, rereplication induced by geminin depletion and oncogene activation requires significant contributions of both Y-Pols and replicative polymerases. These findings offer important mechanistic insights into cancer genomic instability.
authors.affiliation: Laboratory of Molecular Genetics, The Institute for Molecular and Cellular Regulation, Gunma University, Maebashi, Gunma, Japan.
authors.forename: Takayuki
authors.initials: T
authors.lastname: Sekimoto
authors.affiliation: Laboratory of Molecular Genetics, The Institute for Molecular and Cellular Regulation, Gunma University, Maebashi, Gunma, Japan.
authors.forename: Tsukasa
authors.initials: T
authors.lastname: Oda
authors.affiliation: Laboratory of Molecular Genetics, The Institute for Molecular and Cellular Regulation, Gunma University, Maebashi, Gunma, Japan.
authors.forename: Kiminori
authors.initials: K
authors.lastname: Kurashima
authors.affiliation: Faculty of Science, Gakushuin University, Tokyo, Japan.
authors.forename: Fumio
authors.initials: F
authors.lastname: Hanaoka
authors.affiliation: Laboratory of Molecular Genetics, The Institute for Molecular and Cellular Regulation, Gunma University, Maebashi, Gunma, Japan y-taka@gunma-u.ac.jp.
authors.forename: Takayuki
authors.initials: T
authors.lastname: Yamashita
mesh_terms: D045744:Cell Line, Tumor; D019927:Cyclin E / Q000235:genetics / Q000378:metabolism; D004249:DNA Damage; D004260:DNA Repair; D004261:DNA Replication; D004273:DNA, Neoplasm / Q000235:genetics* / Q000378:metabolism; D004259:DNA-Directed DNA Polymerase / Q000235:genetics* / Q000378:metabolism; D064248:Geminin / Q000172:deficiency / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic; D005822:Genetic Vectors; D016678:Genome; D042822:Genomic Instability; D045325:HCT116 Cells; D006801:Humans; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D016086:Lentivirus / Q000235:genetics; D019076:Transgenes
chemical_list: D019927:Cyclin E; D004273:DNA, Neoplasm; C577995:GMNN protein, human; D064248:Geminin; D007527:Isoenzymes; D004259:DNA-Directed DNA Polymerase