Pol η is required for DNA replication during nucleotide deprivation by hydroxyurea

Feraudy, Sébastien de; Limoli, Charles L.; Giedzinski, Erich; Karentz, Deneb; Marti, Thomas; Feeney, Luzviminda; Cleaver, James E.

doi:10.1038/sj.onc.1210385

Cited by 38 publications

(33 citation statements)

References 38 publications

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“…The generation of RPA-primed single-stranded DNA, a result of extensive DNA unwinding at a stalled replication fork, is essential for triggering RAD18-dependent monoubiquitination of PCNA (11,14). Consistent with this model, DNA damage-induced Pol and REV1 focus formation occurs in response to agents that strongly induce PCNA monoubiquitination, such as UV-C irradiation, hydroxyurea, aphidicolin, and cisplatin (8,15,29,39). Treatments which predominantly introduce interstrand cross-links (e.g., MMC) or alternatively, DSBs (e.g., ionizing radiation and camptothecin, a topoisomerase I poison), do not effectively induce PCNA monubiquitination or TLS polymerase focus formation (see Fig.…”

Section: Discussionsupporting

confidence: 54%

Differential Roles for DNA Polymerases Eta, Zeta, and REV1 in Lesion Bypass of Intrastrand versus Interstrand DNA Cross-Links

Hicks¹,

Chute²,

Paulsen³

et al. 2010

Molecular and Cellular Biology

115

164

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Translesion DNA synthesis (TLS) is a process whereby specialized DNA polymerases are recruited to bypass DNA lesions that would otherwise stall high-fidelity polymerases. We provide evidence that TLS across cisplatin intrastrand cross-links is performed by multiple translesion DNA polymerases. First, we determined that PCNA monoubiquitination by RAD18 is necessary for efficient bypass of cisplatin adducts by the TLS polymerases eta (Pol), REV1, and zeta (Pol) based on the observations that depletion of these proteins individually leads to decreased cell survival, cell cycle arrest in S phase, and activation of the DNA damage response. Second, we showed that in addition to PCNA monoubiquitination by RAD18, the Fanconi anemia core complex is also important for recruitment of REV1 to stalled replication forks in cisplatin treated cells. Third, we present evidence that REV1 and Pol are uniquely associated with protection against cisplatin and mitomycin C-induced chromosomal aberrations, and both are necessary for the timely resolution of DNA double-strand breaks associated with repair of DNA interstrand cross-links. Together, our findings indicate that REV1 and Pol facilitate repair of interstrand cross-links independently of PCNA monoubiquitination and Pol, whereas RAD18 plus Pol, REV1, and Pol are all necessary for replicative bypass of cisplatin intrastrand DNA cross-links.Maintenance of genomic integrity involves the activation of cell cycle checkpoints coupled with DNA repair. Despite these sophisticated mechanisms to remove DNA lesions prior to DNA replication, replication forks may inevitably encounter nonrepaired lesions that block high fidelity polymerases, potentially leading to replication fork instability, gaps in replicated DNA, and the generation of DNA double-strand breaks (DSBs). In order to preserve replication fork stability by allowing replication through polymerase blocking lesions, template DNA containing a damaged base or abasic site can be replicated through the actions of specialized translesion DNA synthesis (TLS) polymerases (61). A key event in the regulation of TLS is the monoubiquitination of PCNA, a homotrimeric protein that functions as an auxiliary factor for DNA polymerases (28,31,57,60). The RAD6 (E2)-RAD18 (E3) complex specifically monoubiquitinates PCNA on Lys-164 in response to replication fork stalling. This event is thought to operate as a molecular switch from normal DNA replication to the TLS pathway based on the observations that association of Y-family TLS polymerases with monoubiquitinated PCNA is strengthened through the cooperative binding of one or more ubiquitin-binding domains (UBM or UBZ) plus a PCNA-interacting domain (6,25).Extensive biochemical evidence suggests that replication through a large variety of lesions requires the sequential action of two TLS polymerases (44). The Y-family polymerase eta (Pol) plays a key role in the efficient and error-free bypass of cyclobutane pyrimidine (TT) dimers, one of the major lesions resulting from exposure to UV radiation (...

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Section: Discussionsupporting

confidence: 54%

Differential Roles for DNA Polymerases Eta, Zeta, and REV1 in Lesion Bypass of Intrastrand versus Interstrand DNA Cross-Links

Hicks¹,

Chute²,

Paulsen³

et al. 2010

Molecular and Cellular Biology

115

164

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“…While the former process requires Pol ␣ primase and unphosphorylated RPA-dependent initiation at replication origins and relies heavily on two other processive, high-fidelity polymerases (Pol ␦ and Pol ε) at replication elongation, DNA synthesis in the recovering cells mainly resumes at the arrested replication forks or starts as part of a repair process and therefore may not need the initiation step and may be mediated by nonreplicative but repair-specific polymerases (23,42,51). In support of this idea, several nonreplicative DNA polymerases, including X-family polymerases , , TdT, Y-family polymerase , B-family polymerase , and Rev1, have been found to be involved in the nonhomologous end joining and homologous recombination repair synthesis of DSBs, the major type of DNA breaks induced by collapse of replication forks following HU exposure (12,19,33,42,51). In addition, it has also been shown that the Pol ␣-primase complex is not required for DSB-induced homologous recombination during the mating type switch in budding yeast (50).…”

Section: Discussionmentioning

confidence: 66%

Protein Phosphatase 2A-Dependent Dephosphorylation of Replication Protein A Is Required for the Repair of DNA Breaks Induced by Replication Stress

Feng

Wakeman

Yong

et al. 2009

Molecular and Cellular Biology

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Eukaryotic genomic integrity is safeguarded by cell cycle checkpoints and DNA repair pathways, collectively known as the DNA damage response, wherein replication protein A (RPA) is a key regulator playing multiple critical roles. The genotoxic insult-induced phosphorylation of the 32-kDa subunit of human RPA (RPA32), most notably the ATM/ATR-dependent phosphorylation at T21 and S33, acts to suppress DNA replication and recruit other checkpoint/repair proteins to the DNA lesions. It is not clear, however, how the DNA damageresponsive function of phosphorylated RPA is attenuated and how the replication-associated activity of the unphosphorylated form of RPA is restored when cells start to resume the normal cell cycle. We report here that in cells recovering from hydroxyurea (HU)-induced genotoxic stress, RPA32 is dephosphorylated by the serine/threonine protein phosphatase 2A (PP2A). Interference with PP2A catalytic activity causes persistent RPA32 phosphorylation and increased HU sensitivity. The PP2A catalytic subunit binds to RPA following DNA damage and can dephosphorylate RPA32 in vitro. Cells expressing a RPA32 persistent phosphorylation mimetic exhibit normal checkpoint activation and reenter the cell cycle normally after recovery but display a pronounced defect in the repair of DNA breaks. These data indicate that PP2A-mediated RPA32 dephosphorylation is required for the efficient DNA damage repair.

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“…The 293T human embryonic kidney epithelial cells were purchased from ATCC. The SV40-transformed Pol -deficient XP30RO fibroblasts and the corrected cells (XP30RO ϩ Pol ) were provided by Professor James E. Cleaver (33,34). Cells were cultured in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum (Invitrogen), 100 units/ml of penicillin, and 100 g/ml of streptomycin (ATCC), and incubated at 37°C in 5% CO 2 atmosphere.…”

Section: Methodsmentioning

confidence: 99%

Translesion Synthesis of 8,5′-Cyclopurine-2′-deoxynucleosides by DNA Polymerases η, ι, and ζ

You¹,

Swanson

Dai³

et al. 2013

Journal of Biological Chemistry

145

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Background:The 8,5Ј-cyclopurine-2Ј-deoxynucleosides (cPus) are important types of oxidative DNA damage. Results: cPus exhibit both inhibitory and mutagenic effects on replication; polymerases (Pols) , , and are involved in translesion synthesis of these lesions. Conclusion: Pols , , and cooperatively promote translesion synthesis across cPu lesions. Significance: This work revealed the effects of cPus on DNA replication in mammalian cells.

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Pol η is required for DNA replication during nucleotide deprivation by hydroxyurea

Cited by 38 publications

References 38 publications

Differential Roles for DNA Polymerases Eta, Zeta, and REV1 in Lesion Bypass of Intrastrand versus Interstrand DNA Cross-Links

Differential Roles for DNA Polymerases Eta, Zeta, and REV1 in Lesion Bypass of Intrastrand versus Interstrand DNA Cross-Links

Protein Phosphatase 2A-Dependent Dephosphorylation of Replication Protein A Is Required for the Repair of DNA Breaks Induced by Replication Stress

Translesion Synthesis of 8,5′-Cyclopurine-2′-deoxynucleosides by DNA Polymerases η, ι, and ζ

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