Adduct formation and repair, and translesion DNA synthesis across the adducts in human cells exposed to 3-nitrobenzanthrone

Kawanishi, Michihiro; Fujikawa, Yukichi; Ishii, Hiroshi; Nishida, Hiroshi; Higashigaki, Yuka; Kanno, Takaharu; Matsuda, Tomonari; Takamura-Enya, Takeji; Yagi, Takashi

doi:10.1016/j.mrgentox.2013.03.005

Cited by 20 publications

(61 citation statements)

References 51 publications

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“…60 In this work, in contrast to our results, the C8-dG-ABA adduct predominantly caused G → A mutations (15.0%), followed by G → T (8.8%) and G → C (2.0%) mutations in human skin fibroblast cell line XP2OS(SV). 60 We were surprised by the change in mutational specificity between the two different human cell lines, even though an NER-deficient cell line was used in the published work, whereas we used NER-proficient HEK293T cells. In addition to the difference in cell lines, the DNA sequence context beyond the immediate neighbors of the lesion was dissimilar in this work.…”

Section: Resultscontrasting

confidence: 99%

“…However, when the scaffold contained a C opposite C8-dG-ABA, G → A events were less than 1% as in the single-stranded vector (Table S4L of the Supporting Information and Figure 4B). On the basis of these experiments, we conclude that the presence of C8-dG-ABA in the bubble region of the duplex plasmid, in addition to the different sequence context and the type of cells, has contributed to the array of mutations noted in ref (60). …”

Section: Resultsmentioning

confidence: 66%

“…60 This difference may originate from the differences in the local DNA sequence contexts and the types of cells used. In addition, we used a single-stranded shuttle vector, whereas the reported study was performed in a duplex vector in which the C8-dG-ABA adduct was located in a bubble region.…”

Section: Discussionmentioning

confidence: 99%

“…In addition, we used a single-stranded shuttle vector, whereas the reported study was performed in a duplex vector in which the C8-dG-ABA adduct was located in a bubble region. 60 To determine if the bubble region influenced the types of mutations, we replicated a construct in which C8-dG-ABA was placed in a similar bubble. For this experiment, we used COS-7, a different type of mammalian cell line, because we also wanted to determine if mutational types might vary in a dissimilar mammalian cell line.…”

Section: Discussionmentioning

confidence: 99%

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Mutational Analysis of the C8-Guanine Adduct of the Environmental Carcinogen 3-Nitrobenzanthrone in Human Cells: Critical Roles of DNA Polymerases η and κ and Rev1 in Error-Prone Translesion Synthesis

et al. 2014

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3-Nitrobenzanthrone (3-NBA), a potent mutagen and suspected human carcinogen, is a common environmental pollutant. The genotoxicity of 3-NBA has been associated with its ability to form DNA adducts, including N-(2′-deoxyguanosin-8-yl)-3-aminobenzanthrone (C8-dG-ABA). To investigate the molecular mechanism of C8-dG-ABA mutagenesis in human cells, we have replicated a plasmid containing a single C8-dG-ABA in human embryonic kidney 293T (HEK293T) cells, which yielded 14% mutant progeny. The major types of mutations induced by C8-dG-ABA were G → T > G → A > G → C. siRNA knockdown of the translesion synthesis (TLS) DNA polymerases (pols) in HEK293T cells indicated that pol η, pol κ, pol ι, pol ζ, and Rev1 each have a role in replication across this adduct. The extent of TLS was reduced with each pol knockdown, but the largest decrease (of ∼55% reduction) in the level of TLS occurred in cells with knockdown of pol ζ. Pol η and pol κ were considered the major contributors of the mutagenic TLS, because the mutation frequency (MF) decreased by 70%, when these pols were simultaneously knocked down. Rev1 also is important for mutagenesis, as reflected by the 60% reduction in MF upon Rev1 knockdown, but it probably plays a noncatalytic role by physically interacting with the other two Y-family pols. In contrast, pol ζ appeared to be involved in the error-free bypass of the lesion, because MF increased by 60% in pol ζ knockdown cells. These results provide important mechanistic insight into the bypass of the C8-dG-ABA adduct.

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Section: Resultscontrasting

confidence: 99%

Section: Resultsmentioning

confidence: 66%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Mutational Analysis of the C8-Guanine Adduct of the Environmental Carcinogen 3-Nitrobenzanthrone in Human Cells: Critical Roles of DNA Polymerases η and κ and Rev1 in Error-Prone Translesion Synthesis

et al. 2014

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“…This cellular TLS assay is superior to the cell free TLS assay in a test tube using adducted oligonucleotide, extension primer and puriˆed TLS polymerases because the in vitro test tube assay is less re‰ective of the cellular process. A project to measure TLS e‹ciencies and mutation frequencies of various DNA adducts formed by 3-nitrobenzanthrone, a carcinogenic nitro-PAH from diesel exhaust, is in progress (62).…”

Section: Prospects For New Shuttle Vector Development To Study Tlsmentioning

confidence: 99%

The Achievement of Shuttle Vector Techniques in Mammalian Cell Mutation Research

Yagi¹

2013

Genes and Environment

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Shuttle vector plasmids have made major contributions to the advancement of mutation research for the last two decades. I have been involved in the development of shuttle vector plasmids and their experimental protocols, and herein provide a chronicle of shuttle vector studies. In the late 1960s, in vitro mammalian cell culture became a common technique in cell biology. Geneticists established a method to detect gene mutation as 8-azaguanine or 6-thioguanine resistance after cells had been exposed to radiation or chemicals. The resistance of the cells was found to be due to a mutation of the hypoxanthine-guanine phosphoribosyl transferase gene; however, which base was changed in the gene remained di‹cult to determine until DNA sequencing techniques were developed. After the Sanger method of DNA sequencing became available to geneticists, the next issue was how to identify numerous base changes in mutants easily and rapidly. A breakthrough on this issue was the use of the shuttle vector plasmid pZ189, which was developed for practical applications by Seidman and colleagues. Along with the development of new molecular biology techniques such as polymerase chain reaction and automatic DNA sequencing, pZ189 has been modiˆed to several forms as adaptations to the new techniques. These shuttle vector plasmids remain useful tools to reveal what types of mutations are induced by newly identiˆed mutagens at the DNA sequence level. This article describes some of the contents of my JEMS award lecture in 2011.

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Mechanism of Error‐Free Bypass of the Environmental Carcinogen N‐(2′‐Deoxyguanosin‐8‐yl)‐3‐aminobenzanthrone Adduct by Human DNA Polymerase η

et al. 2016

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The environmental pollutant 3‐nitrobenzanthrone produces bulky aminobenzanthrone (ABA) DNA adducts with both guanine and adenine nucleobases. A major product occurs at the C8 position of guanine (C8‐dG‐ABA). These adducts present a strong block to replicative polymerases but, remarkably, can be bypassed in a largely error‐free manner by the human Y‐family polymerase η (hPol η). Here, we report the crystal structure of a ternary Pol⋅DNA⋅dCTP complex between a C8‐dG‐ABA‐containing template:primer duplex and hPol η. The complex was captured at the insertion stage and provides crucial insight into the mechanism of error‐free bypass of this bulky lesion. Specifically, bypass involves accommodation of the ABA moiety inside a hydrophobic cleft to the side of the enzyme active site and formation of an intra‐nucleotide hydrogen bond between the phosphate and ABA amino moiety, allowing the adducted guanine to form a standard Watson–Crick pair with the incoming dCTP.

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Adduct formation and repair, and translesion DNA synthesis across the adducts in human cells exposed to 3-nitrobenzanthrone

Cited by 20 publications

References 51 publications

Mutational Analysis of the C8-Guanine Adduct of the Environmental Carcinogen 3-Nitrobenzanthrone in Human Cells: Critical Roles of DNA Polymerases η and κ and Rev1 in Error-Prone Translesion Synthesis

Mutational Analysis of the C8-Guanine Adduct of the Environmental Carcinogen 3-Nitrobenzanthrone in Human Cells: Critical Roles of DNA Polymerases η and κ and Rev1 in Error-Prone Translesion Synthesis

The Achievement of Shuttle Vector Techniques in Mammalian Cell Mutation Research

Mechanism of Error‐Free Bypass of the Environmental Carcinogen N‐(2′‐Deoxyguanosin‐8‐yl)‐3‐aminobenzanthrone Adduct by Human DNA Polymerase η

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