Translesional DNA Synthesis through a C8-Guanyl Adduct of 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) in Vitro

Fukuda, Hirokazu; Takamura-Enya, Takeji; Masuda, Yuji; Nohmi, Takehiko; Seki, Chiho; Kamiya, Kenji; Sügimura, Takashi; Masutani, Chikahide; Hanaoka, Fumio; Nakagama, Hitoshi

doi:10.1074/jbc.m109.037259

Cited by 11 publications

(2 citation statements)

References 43 publications

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“…Molecular modeling and in vitro studies suggest that C8-PhIP-dG adducts block replicative polymerases, enhance the infidelity of replication and may engage error-prone translesion synthesis (21–23). Interference with the DNA replication machinery can trigger a cellular stress response, referred to as replication stress.…”

Section: Introductionmentioning

confidence: 99%

DNA damage response curtails detrimental replication stress and chromosomal instability induced by the dietary carcinogen PhIP

Mimmler¹,

Schmidt²,

Kraus³

et al. 2016

Nucleic Acids Research

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PhIP is an abundant heterocyclic aromatic amine (HCA) and important dietary carcinogen. Following metabolic activation, PhIP causes bulky DNA lesions at the C8-position of guanine. Although C8-PhIP-dG adducts are mutagenic, their interference with the DNA replication machinery and the elicited DNA damage response (DDR) have not yet been studied. Here, we analyzed PhIP-triggered replicative stress and elucidated the role of the apical DDR kinases ATR, ATM and DNA-PKcs in the cellular defense response. First, we demonstrate that PhIP induced C8-PhIP-dG adducts and DNA strand breaks. This stimulated ATR-CHK1 signaling, phosphorylation of histone 2AX and the formation of RPA foci. In proliferating cells, PhIP treatment increased the frequency of stalled replication forks and reduced fork speed. Inhibition of ATR in the presence of PhIP-induced DNA damage strongly promoted the formation of DNA double-strand breaks, activation of the ATM-CHK2 pathway and hyperphosphorylation of RPA. The abrogation of ATR signaling potentiated the cell death response and enhanced chromosomal aberrations after PhIP treatment, while ATM and DNA-PK inhibition had only marginal effects. These results strongly support the notion that ATR plays a key role in the defense against cancer formation induced by PhIP and related HCAs.

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Section: Introductionmentioning

confidence: 99%

DNA damage response curtails detrimental replication stress and chromosomal instability induced by the dietary carcinogen PhIP

Mimmler¹,

Schmidt²,

Kraus³

et al. 2016

Nucleic Acids Research

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“…Whereas the eukaryotic REV1 protein shares these structural features, its role in TLS is unique. First, Rev1 is a deoxycytidyl transferase that is very specific for incorporating a C opposite G, abasic sites and damaged bases (5)(6)(7)(8)(9)(10)(11). This is in contrast to all other known DNA polymerases that utilize all four nucleotides for DNA synthesis.…”

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confidence: 98%

The Non-canonical Protein Binding Site at the Monomer-Monomer Interface of Yeast Proliferating Cell Nuclear Antigen (PCNA) Regulates the Rev1-PCNA Interaction and Polζ/Rev1-dependent Translesion DNA Synthesis

Sharma¹,

Kochenova

Shcherbakova

2011

Journal of Biological Chemistry

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Rev1 and DNA polymerase (Pol) are involved in the tolerance of DNA damage by translesion synthesis (TLS). The proliferating cell nuclear antigen (PCNA), the auxiliary factor of nuclear DNA polymerases, plays an important role in regulating the access of TLS polymerases to the primer terminus. Both Rev1 and Pol lack the conserved hydrophobic motif that is used by many proteins for the interaction with PCNA at its interdomain connector loop. We have previously reported that the interaction of yeast Pol with PCNA occurs at an unusual site near the monomer-monomer interface of the trimeric PCNA. Using GST pull-down assays, PCNA-coupled affinity beads pulldown and gel filtration chromatography, we show that the same region is required for the physical interaction of PCNA with the polymerase-associated domain (PAD) of Rev1. The interaction is disrupted by the pol30-113 mutation that results in a double amino acid substitution at the monomer-monomer interface of PCNA. Genetic analysis of the epistatic relationship of the pol30-113 mutation with an array of DNA repair and damage tolerance mutations indicated that PCNA-113 is specifically defective in the Rev1/Pol-dependent TLS pathway. Taken together, the data suggest that Pol and Rev1 are unique among PCNA-interacting proteins in using the novel binding site near the intermolecular interface of PCNA. The new mode of Rev1-PCNA binding described here suggests a mechanism by which Rev1 adopts a catalytically inactive configuration at the replication fork.Cellular DNA is continuously attacked by endogenous and exogenous agents that damage the bases and the DNA backbone. Damage that is not repaired prior to the S phase of cell cycle can block the replication machinery, because most lesions cannot be accommodated in the highly selective active site of replicative DNA polymerases (1, 2). Replication stalling activates several damage tolerance mechanisms that help bypass the damage in either an accurate or mutagenic manner. Translesion DNA synthesis (TLS) 3 is an important damage tolerance pathway, in which specialized DNA polymerases are recruited to synthesize DNA through template lesions (3). Structural studies showed that TLS polymerases have a more open active site that allows them to accommodate a variety of DNA lesions and catalyze polymerization on damaged templates (4). The accuracy of TLS can vary depending on the particular lesion and the DNA polymerases involved. It is, however, an inherently mutagenic process, because the damage can alter the coding properties of the bases, and because TLS polymerases generally have low fidelity (2).In human cells, TLS polymerases include Y family enzymes Pol, Pol, Pol, and REV1, and the B family enzyme Pol. Although the Y family polymerases share little amino acid sequence similarity with the classical DNA polymerases, they have a similar overall "right-hand" architecture with the "palm," "thumb," and "fingers" domains. The thumb and fingers domains of the Y family enzymes, however, are short and do not make as many contacts wi...

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Catalytic and non‐catalytic roles of DNA polymerase κ in the protection of human cells against genotoxic stresses

Kanemaru

Suzuki

Niimi

et al. 2015

Environ and Mol Mutagen

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DNA polymerase κ (Pol κ) is a specialized DNA polymerase involved in translesion DNA synthesis. Although its bypass activities across lesions are well characterized in biochemistry, its cellular protective roles against genotoxic insults are still elusive. To better understand the in vivo protective roles, we have established a human cell line deficient in the expression of Pol κ (KO) and another expressing catalytically dead Pol κ (CD), to examine the cytotoxic sensitivity to 11 genotoxins including ultraviolet C light (UV). These cell lines were established in a genetic background of Nalm‐6‐MSH+, a human lymphoblastic cell line that has high efficiency for gene targeting, and functional p53 and mismatch repair activities. We classified the genotoxins into four groups. Group 1 includes benzo[a]pyrene diolepoxide, mitomycin C, and bleomycin, where the sensitivity was equally higher in KO and CD than in the cell line expressing wild‐type Pol κ (WT). Group 2 includes hydrogen peroxide and menadione, where hypersensitivity was observed only in KO. Group 3 includes methyl methanesulfonate and ethyl methanesulfonate, where hypersensitivity was observed only in CD. Group 4 includes UV and three chemicals, where the chemicals exhibited similar cytotoxicity to all three cell lines. The results suggest that Pol κ not only protects cells from genotoxic DNA lesions via DNA polymerase activities, but also contributes to genome integrity by acting as a non‐catalytic protein against oxidative damage caused by hydrogen peroxide and menadione. The non‐catalytic roles of Pol κ in protection against oxidative damage by hydrogen peroxide are discussed. Environ. Mol. Mutagen. 56:650–662, 2015. © 2015 Wiley Periodicals, Inc.

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Translesional DNA Synthesis through a C8-Guanyl Adduct of 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) in Vitro

Cited by 11 publications

References 43 publications

DNA damage response curtails detrimental replication stress and chromosomal instability induced by the dietary carcinogen PhIP

DNA damage response curtails detrimental replication stress and chromosomal instability induced by the dietary carcinogen PhIP

The Non-canonical Protein Binding Site at the Monomer-Monomer Interface of Yeast Proliferating Cell Nuclear Antigen (PCNA) Regulates the Rev1-PCNA Interaction and Polζ/Rev1-dependent Translesion DNA Synthesis

Catalytic and non‐catalytic roles of DNA polymerase κ in the protection of human cells against genotoxic stresses

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