The influence of repair pathways on the cytotoxicity and mutagenicity induced by the pyridyloxobutylation pathway of tobacco-specific nitrosamines

Li Li, Joana Perdigao, Anthony Pegg, Yanbin Lao, Stephen S. Hecht, Bruce R. Lindgren, Joyce T. Reardon, Aziz Sancar, Elizabeth V. Wattenberg, Lisa A. Peterson

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Tobacco-specific nitrosamines, 4-(methylnitrosamino)-1-(3-pyridyl)-1- butanone and N′-nitrosonornicotine, are considered to be human carcinogens. Both compounds are metabolized to pyridyloxobutylating intermediates that react with DNA to form adducts such as 7-[4-(3-pyridyl)-4- oxobut-1-yl]guanine, O2-[4-(3-pyridyl)-4-oxobut-1-yl]cytosine, O 2-[4-(3-pyridyl)-4-oxobut-1-yl]-2′-deoxythymidine (O 2-pobdT), O6-[4-(3-pyridyl)-4-oxobut-1-yl]-2′- deoxyguanosine (O6-pobdG), and 4-hydroxy-1-(3-pyridyl)-1-butanone- releasing adducts. The role of specific DNA adducts in the overall genotoxic activity of the pyridyloxobutylation pathway is not known. One adduct, O 6-pobdG, is mutagenic. To characterize the mutagenic and cytotoxic properties of pyridyloxobutyl DNA adducts, the impact of DNA repair pathways on the cytotoxic and mutagenic properties of the model pyridyloxobutylating agent, 4-(acetoxymethylnitrosamino)-1-(3-pyridyl)-1-butanone (NNKOAc), was investigated in Chinese hamster ovary cell lines proficient or deficient in O 6-alkylguanine DNA alkyltransferase (AGT), deficient in both AGT and base excision repair (BER), or deficient in both AGT and nucleotide excision repair (NER). The repair of the four pyridyloxobutyl DNA adducts was determined in the same cell lines via sensitive LC-MS/MS methods. NNKOAc was more cytotoxic in the cell lines lacking AGT, BER, and NER repair pathways. It also induced more mutations in the hprt gene in the BER- and NER-deficient cell lines. However, AGT expression did not influence NNKOAc's mutagenicity despite efficient repair of O6-pobdG. Analysis of the hprt mutational spectra indicated that NNKOAc primarily caused point mutations at AT base pairs. GC to AT transition mutations were a minor contributor to the overall mutation spectrum, providing a rationale for the observation that AGT does not protect against the overall mutagenic properties of NNKOAc in this model system. The only adduct affected by the absence of effective NER was O2-pobdT. Slower repair of O2-pobdT in NER-deficient cells was associated with increased AT to TA transversion mutations, supporting the hypothesis that these mutations are caused by O2-pobdT. Together, these data support a hypothesis that the pyridyloxobutylation pathway generates multiple mutagenic and toxic adducts.

Original languageEnglish (US)
Pages (from-to)1464-1472
Number of pages9
JournalChemical Research in Toxicology
Volume22
Issue number8
DOIs
StatePublished - Aug 17 2009

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Nitrosamines
Tobacco
Cytotoxicity
DNA Repair
Repair
DNA Adducts
Mutation
Nucleotides
Cell Line
N'-nitrosonornicotine
Cells
O(6)-Methylguanine-DNA Methyltransferase
Deoxyguanosine
Poisons
Cytosine
Guanine
Cricetulus
Point Mutation
Base Pairing
Carcinogens

All Science Journal Classification (ASJC) codes

  • Toxicology

Cite this

Li, Li ; Perdigao, Joana ; Pegg, Anthony ; Lao, Yanbin ; Hecht, Stephen S. ; Lindgren, Bruce R. ; Reardon, Joyce T. ; Sancar, Aziz ; Wattenberg, Elizabeth V. ; Peterson, Lisa A. / The influence of repair pathways on the cytotoxicity and mutagenicity induced by the pyridyloxobutylation pathway of tobacco-specific nitrosamines. In: Chemical Research in Toxicology. 2009 ; Vol. 22, No. 8. pp. 1464-1472.
@article{246931cbc24b49499c4bd014cc618f0a,
title = "The influence of repair pathways on the cytotoxicity and mutagenicity induced by the pyridyloxobutylation pathway of tobacco-specific nitrosamines",
abstract = "Tobacco-specific nitrosamines, 4-(methylnitrosamino)-1-(3-pyridyl)-1- butanone and N′-nitrosonornicotine, are considered to be human carcinogens. Both compounds are metabolized to pyridyloxobutylating intermediates that react with DNA to form adducts such as 7-[4-(3-pyridyl)-4- oxobut-1-yl]guanine, O2-[4-(3-pyridyl)-4-oxobut-1-yl]cytosine, O 2-[4-(3-pyridyl)-4-oxobut-1-yl]-2′-deoxythymidine (O 2-pobdT), O6-[4-(3-pyridyl)-4-oxobut-1-yl]-2′- deoxyguanosine (O6-pobdG), and 4-hydroxy-1-(3-pyridyl)-1-butanone- releasing adducts. The role of specific DNA adducts in the overall genotoxic activity of the pyridyloxobutylation pathway is not known. One adduct, O 6-pobdG, is mutagenic. To characterize the mutagenic and cytotoxic properties of pyridyloxobutyl DNA adducts, the impact of DNA repair pathways on the cytotoxic and mutagenic properties of the model pyridyloxobutylating agent, 4-(acetoxymethylnitrosamino)-1-(3-pyridyl)-1-butanone (NNKOAc), was investigated in Chinese hamster ovary cell lines proficient or deficient in O 6-alkylguanine DNA alkyltransferase (AGT), deficient in both AGT and base excision repair (BER), or deficient in both AGT and nucleotide excision repair (NER). The repair of the four pyridyloxobutyl DNA adducts was determined in the same cell lines via sensitive LC-MS/MS methods. NNKOAc was more cytotoxic in the cell lines lacking AGT, BER, and NER repair pathways. It also induced more mutations in the hprt gene in the BER- and NER-deficient cell lines. However, AGT expression did not influence NNKOAc's mutagenicity despite efficient repair of O6-pobdG. Analysis of the hprt mutational spectra indicated that NNKOAc primarily caused point mutations at AT base pairs. GC to AT transition mutations were a minor contributor to the overall mutation spectrum, providing a rationale for the observation that AGT does not protect against the overall mutagenic properties of NNKOAc in this model system. The only adduct affected by the absence of effective NER was O2-pobdT. Slower repair of O2-pobdT in NER-deficient cells was associated with increased AT to TA transversion mutations, supporting the hypothesis that these mutations are caused by O2-pobdT. Together, these data support a hypothesis that the pyridyloxobutylation pathway generates multiple mutagenic and toxic adducts.",
author = "Li Li and Joana Perdigao and Anthony Pegg and Yanbin Lao and Hecht, {Stephen S.} and Lindgren, {Bruce R.} and Reardon, {Joyce T.} and Aziz Sancar and Wattenberg, {Elizabeth V.} and Peterson, {Lisa A.}",
year = "2009",
month = "8",
day = "17",
doi = "10.1021/tx9001572",
language = "English (US)",
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}

Li, L, Perdigao, J, Pegg, A, Lao, Y, Hecht, SS, Lindgren, BR, Reardon, JT, Sancar, A, Wattenberg, EV & Peterson, LA 2009, 'The influence of repair pathways on the cytotoxicity and mutagenicity induced by the pyridyloxobutylation pathway of tobacco-specific nitrosamines', Chemical Research in Toxicology, vol. 22, no. 8, pp. 1464-1472. https://doi.org/10.1021/tx9001572

The influence of repair pathways on the cytotoxicity and mutagenicity induced by the pyridyloxobutylation pathway of tobacco-specific nitrosamines. / Li, Li; Perdigao, Joana; Pegg, Anthony; Lao, Yanbin; Hecht, Stephen S.; Lindgren, Bruce R.; Reardon, Joyce T.; Sancar, Aziz; Wattenberg, Elizabeth V.; Peterson, Lisa A.

In: Chemical Research in Toxicology, Vol. 22, No. 8, 17.08.2009, p. 1464-1472.

Research output: Contribution to journalArticle

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T1 - The influence of repair pathways on the cytotoxicity and mutagenicity induced by the pyridyloxobutylation pathway of tobacco-specific nitrosamines

AU - Li, Li

AU - Perdigao, Joana

AU - Pegg, Anthony

AU - Lao, Yanbin

AU - Hecht, Stephen S.

AU - Lindgren, Bruce R.

AU - Reardon, Joyce T.

AU - Sancar, Aziz

AU - Wattenberg, Elizabeth V.

AU - Peterson, Lisa A.

PY - 2009/8/17

Y1 - 2009/8/17

N2 - Tobacco-specific nitrosamines, 4-(methylnitrosamino)-1-(3-pyridyl)-1- butanone and N′-nitrosonornicotine, are considered to be human carcinogens. Both compounds are metabolized to pyridyloxobutylating intermediates that react with DNA to form adducts such as 7-[4-(3-pyridyl)-4- oxobut-1-yl]guanine, O2-[4-(3-pyridyl)-4-oxobut-1-yl]cytosine, O 2-[4-(3-pyridyl)-4-oxobut-1-yl]-2′-deoxythymidine (O 2-pobdT), O6-[4-(3-pyridyl)-4-oxobut-1-yl]-2′- deoxyguanosine (O6-pobdG), and 4-hydroxy-1-(3-pyridyl)-1-butanone- releasing adducts. The role of specific DNA adducts in the overall genotoxic activity of the pyridyloxobutylation pathway is not known. One adduct, O 6-pobdG, is mutagenic. To characterize the mutagenic and cytotoxic properties of pyridyloxobutyl DNA adducts, the impact of DNA repair pathways on the cytotoxic and mutagenic properties of the model pyridyloxobutylating agent, 4-(acetoxymethylnitrosamino)-1-(3-pyridyl)-1-butanone (NNKOAc), was investigated in Chinese hamster ovary cell lines proficient or deficient in O 6-alkylguanine DNA alkyltransferase (AGT), deficient in both AGT and base excision repair (BER), or deficient in both AGT and nucleotide excision repair (NER). The repair of the four pyridyloxobutyl DNA adducts was determined in the same cell lines via sensitive LC-MS/MS methods. NNKOAc was more cytotoxic in the cell lines lacking AGT, BER, and NER repair pathways. It also induced more mutations in the hprt gene in the BER- and NER-deficient cell lines. However, AGT expression did not influence NNKOAc's mutagenicity despite efficient repair of O6-pobdG. Analysis of the hprt mutational spectra indicated that NNKOAc primarily caused point mutations at AT base pairs. GC to AT transition mutations were a minor contributor to the overall mutation spectrum, providing a rationale for the observation that AGT does not protect against the overall mutagenic properties of NNKOAc in this model system. The only adduct affected by the absence of effective NER was O2-pobdT. Slower repair of O2-pobdT in NER-deficient cells was associated with increased AT to TA transversion mutations, supporting the hypothesis that these mutations are caused by O2-pobdT. Together, these data support a hypothesis that the pyridyloxobutylation pathway generates multiple mutagenic and toxic adducts.

AB - Tobacco-specific nitrosamines, 4-(methylnitrosamino)-1-(3-pyridyl)-1- butanone and N′-nitrosonornicotine, are considered to be human carcinogens. Both compounds are metabolized to pyridyloxobutylating intermediates that react with DNA to form adducts such as 7-[4-(3-pyridyl)-4- oxobut-1-yl]guanine, O2-[4-(3-pyridyl)-4-oxobut-1-yl]cytosine, O 2-[4-(3-pyridyl)-4-oxobut-1-yl]-2′-deoxythymidine (O 2-pobdT), O6-[4-(3-pyridyl)-4-oxobut-1-yl]-2′- deoxyguanosine (O6-pobdG), and 4-hydroxy-1-(3-pyridyl)-1-butanone- releasing adducts. The role of specific DNA adducts in the overall genotoxic activity of the pyridyloxobutylation pathway is not known. One adduct, O 6-pobdG, is mutagenic. To characterize the mutagenic and cytotoxic properties of pyridyloxobutyl DNA adducts, the impact of DNA repair pathways on the cytotoxic and mutagenic properties of the model pyridyloxobutylating agent, 4-(acetoxymethylnitrosamino)-1-(3-pyridyl)-1-butanone (NNKOAc), was investigated in Chinese hamster ovary cell lines proficient or deficient in O 6-alkylguanine DNA alkyltransferase (AGT), deficient in both AGT and base excision repair (BER), or deficient in both AGT and nucleotide excision repair (NER). The repair of the four pyridyloxobutyl DNA adducts was determined in the same cell lines via sensitive LC-MS/MS methods. NNKOAc was more cytotoxic in the cell lines lacking AGT, BER, and NER repair pathways. It also induced more mutations in the hprt gene in the BER- and NER-deficient cell lines. However, AGT expression did not influence NNKOAc's mutagenicity despite efficient repair of O6-pobdG. Analysis of the hprt mutational spectra indicated that NNKOAc primarily caused point mutations at AT base pairs. GC to AT transition mutations were a minor contributor to the overall mutation spectrum, providing a rationale for the observation that AGT does not protect against the overall mutagenic properties of NNKOAc in this model system. The only adduct affected by the absence of effective NER was O2-pobdT. Slower repair of O2-pobdT in NER-deficient cells was associated with increased AT to TA transversion mutations, supporting the hypothesis that these mutations are caused by O2-pobdT. Together, these data support a hypothesis that the pyridyloxobutylation pathway generates multiple mutagenic and toxic adducts.

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