Mutagenicity of a unique apurinic/apyrimidinic site in mammalian cells

Gentil, A.; Cabral‐Neto, Januário B.; Mariage‐Samson, Régine; Margot, A.; Imbach, J.‐L.; Rayner, Bernard; Sarasin, Alain

doi:10.1016/0022-2836(92)90513-j

Cited by 84 publications

(53 citation statements)

References 12 publications

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“…The similar results obtained by using vectors unable or capable of autonomous replication strengthen the physiological relevance of an assay system based on transient transfection with a nonreplicating plasmid to the study of TLR in mammalians cells. It should be mentioned that in two other studies based on shuttle vectors each carrying an abasic lesion at a specific site, there was no apparent preference for a particular nucleotide opposite the abasic site (48,49). The reason for the discrepancy between the various studies is not clear.…”

Section: Discussionmentioning

confidence: 94%

Quantitative measurement of translesion replication in human cells: Evidence for bypass of abasic sites by a replicative DNA polymerase

Avkin

Adar

Blander

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

109

104

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Mutations in oncogenes and tumor suppressor genes are critical in the development of cancer. A major pathway for the formation of mutations is the replication of unrepaired DNA lesions. To better understand the mechanism of translesion replication (TLR) in mammals, a quantitative assay for TLR in cultured cells was developed. The assay is based on the transient transfection of cultured cells with a gapped plasmid, carrying a site-specific lesion in the gap region. Filling in of the gap by TLR is assayed in a subsequent bioassay, by the ability of the plasmid extracted from the cells, to transform an Escherichia coli indicator strain. Using this method it was found that TLR through a synthetic abasic site in the adenocarcinoma H1299, the osteogenic sarcoma Saos-2, the prostate carcinoma PC3, and the hepatoma Hep3B cell lines occurred with efficiencies of 92 ؎ 6%, 32 ؎ 2%, 72 ؎ 4%, and 26 ؎ 3%, respectively. DNA sequence analysis showed that 85% of the bypass events in H1299 cells involved insertion of dAMP opposite the synthetic abasic site. Addition of aphidicolin, an inhibitor of DNA polymerases ␣, ␦, and , caused a 4.4-fold inhibition of bypass. Analysis of two XP-V cell lines, defective in DNA polymerase , showed bypass of 89%, indicating that polymerase is not essential for bypass of abasic sites. These results suggest that in human cells bypass of abasic sites does not require the bypass-specific DNA polymerase , but it does require at least one of the replicative DNA polymerases, ␣, ␦, or . The quantitative TLR assay is expected to be useful in the molecular analysis of lesion bypass in a large variety of cultured mammalian cells.T he formation of mutations is a key event in several important biological phenomena, most notably cancer and evolution. A major mechanism for the formation of mutations is the replication of DNA lesions that have escaped DNA repair (1). For many years the molecular mechanism of this process, termed translesion replication (TLR), translesion synthesis, or lesion bypass, was not clear. Recently it was discovered that the main enzyme responsible for carrying out lesion bypass is a novel type of DNA polymerase, specialized for replicating across damaged sites in DNA. Such DNA polymerases were found in organisms ranging from Escherichia coli (DNA polymerase V; refs. 2 and 3) to humans (e.g., DNA polymerase ; refs. 4 and 5). These novel DNA polymerases comprise the new Y superfamily of DNA polymerases (6). Remarkably, humans contain four members of the Y family: DNA polymerases (4, 5), (7,8), and (9-12) and hREV1, which has dCMP transferase activity (13,14). In addition, humans are likely to have an additional polymerase involved in TLR, polymerase , product of the hREV3 and hREV7 genes (refs. 15 and 16; for review see refs. 17-23).Despite the importance of TLR mechanisms in understanding the role of mutagenesis in cancer, and despite the progress made with the discovery of Y family DNA polymerases, little is known about the mechanism of lesion bypass in mammalian cells. This is partl...

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Section: Discussionmentioning

confidence: 94%

Quantitative measurement of translesion replication in human cells: Evidence for bypass of abasic sites by a replicative DNA polymerase

Avkin

Adar

Blander

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

109

104

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“…Such AP sites are cytotoxic and mutagenic, and must be further processed [3]. Some DNA glycosylases also have an associated AP lyase activity that cleaves the phosphodiester bond 3h to the AP site ( β-lyase).…”

Section: Dna N-glycosylases and The Ber Pathwaymentioning

confidence: 99%

DNA glycosylases in the base excision repair of DNA

Krokan

Standal

Slupphaug

1997

Biochemical Journal

766

566

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A wide range of cytotoxic and mutagenic DNA bases are removed by different DNA glycosylases, which initiate the base excision repair pathway. DNA glycosylases cleave the Nglycosylic bond between the target base and deoxyribose, thus releasing a free base and leaving an apurinic\apyrimidinic (AP) site. In addition, several DNA glycosylases are bifunctional, since they also display a lyase activity that cleaves the phosphodiester backbone 3h to the AP site generated by the glycosylase activity. Structural data and sequence comparisons have identified common features among many of the DNA glycosylases. Their active sites have a structure that can only bind extrahelical target bases, as observed in the crystal structure of human uracil-DNA glycosylase in a complex with double-stranded DNA. Nucleotide flipping is apparently actively facilitated by the enzyme. With bacteriophage T4 endonuclease V, a pyrimidine-

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“…If so, the enzyme would generate an apurinic site at G1 or G2 and this lesion has to be bypassed by TLS with a preferential insertion of dCMP opposite the apurinic site to restore the original G:C pair. However, it is generally agreed that other nucleotides, especially dATP, are preferably inserted opposite the lesion in mammalian cells (26)(27)(28). Thus, the base excision repair mechanism can not explain the repair with such a high fidelity.…”

Section: Icl Repair Mechanismmentioning

confidence: 99%

Replication-Coupled Repair of Crotonaldehyde/Acetaldehyde-Induced Guanine−Guanine Interstrand Cross-Links and Their Mutagenicity

et al. 2006

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The repair of acetaldehyde/crotonaldehyde-induced guanine(N 2 )-guanine(N 2 ) interstrand cross-links (ICLs), 3-(2-deoxyribos-1-yl)-5,6,7,8-(N 2 -deoxyguanosyl)-6(R or S)-methyl-pyrimido [1,2-α] purine-10(3H)one (Scheme 1), was studied using a shuttle plasmid bearing a site-specific ICL. Since the authentic ICLs can revert to monoadducts, a chemically stable model ICL, 1,3-bis(2′-deoxyguanos-N 2 -yl)butane derivative, was also employed to probe the ICL repair mechanism. Since the removal of ICL depends on the nucleotide excision repair (NER) mechanism in Escherichia coli, the plasmid bearing the model ICL failed to yield transformants in NER-deficient host cells, proving the stability of this ICL in cells. The authentic ICLs yielded transformants in the NERdeficient hosts; therefore, these transformants are produced by plasmid bearing spontaneously reverted monoadducts. In contrast, in NER-deficient human cells, the model ICL was removed by an NER-independent repair pathway, which is unique to higher eukaryotes. This repair did not associate with a transcriptional event, but with replication. The analysis of repaired molecules revealed that the authentic and model ICLs were repaired mostly (>94 %) in an error-free manner in both hosts. The major mutations observed were G→T transversions targeting the cross-linked dG located in the lagging strand template. These results support one of the current models for the mammalian NER-independent ICL repair mechanism, in which a DNA endonuclease(s) unhooks an ICL from the leading strand template at a stalled replication fork site by incising on both sides of the ICL and then translesion synthesis is conducted across the "half-excised" ICL attached to the lagging strand template to restore DNA synthesis.Acetaldehyde is ubiquitous in the human environment (1). It is the major metabolite of ethanol and occurs widely in the diet. It is a common environmental pollutant and one of the most prevalent carcinogens in cigarette smoke (2). Crotonaldehyde also occurs in the human environment and in cigarette smoke, and is a product of lipid peroxidation (3-5). These chemicals are genotoxic carcinogens (1,3). DNA adduct formation is considered to be critical in their mechanisms of carcinogenesis. They react with DNA to form Interstrand cross-links (ICLs) 1 in 5′CpG sequence, as illustrated in Scheme 1, and other adducts (6). Similar ICLs † This study was supported by U.S. Public Health Service Grants ES11297, CA76163 and CA47995. ICLs are one of the most serious types of damage to DNA since they connect two strands of DNA covalently and thereby inhibit DNA strand separation required for various important processes such as replication, transcription and recombination. In Escherichia coli, the combined action of nucleotide excision repair (NER) and recombination or translesion synthesis (TLS) has been proposed for the repair of ICLs (reviewed in 11), in which NER incises one strand on both sides of an ICL to unhook from one strand and then the gap generated is filled by recomb...

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Mutagenicity of a unique apurinic/apyrimidinic site in mammalian cells

Cited by 84 publications

References 12 publications

Quantitative measurement of translesion replication in human cells: Evidence for bypass of abasic sites by a replicative DNA polymerase

Quantitative measurement of translesion replication in human cells: Evidence for bypass of abasic sites by a replicative DNA polymerase

DNA glycosylases in the base excision repair of DNA

Replication-Coupled Repair of Crotonaldehyde/Acetaldehyde-Induced Guanine−Guanine Interstrand Cross-Links and Their Mutagenicity

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