Source:http://linkedlifedata.com/resource/pubmed/id/11676538
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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
3
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| pubmed:dateCreated |
2001-10-25
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| pubmed:abstractText |
The nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) is an important tobacco-specific carcinogen associated with lung cancer. Its complex enzymatic activation, leading to methyl and pyridyloxobutyl (POB)-modified DNA, makes DNA damage difficult to characterize and quantify. Therefore, we use the NNK analogue 4-[(acetoxymethyl)nitrosamino]-1-(3-pyridyl)-1-butanone (NNKOAc) to induce damage in genomic DNA, and to map the sites and frequency of adducts at nucleotide resolution using ligation-mediated polymerase chain reaction and terminal transferase-dependent polymerase chain reactions (LMPCR and TDPCR). NNKOAc induced single-strand breaks in a concentration-dependent manner. Post-alkylation treatments, including hot piperidine or digestion with the enzymes Escherichia coli 3-methyladenine-DNA glycosylase II, formamidopyrimidine-DNA glycosylase, Escherichia coli endonuclease III, or phage T4 UV endonuclease V did not increase the level of DNA breaks in NNKOAc-treated DNA. Detection of DNA damage using LMPCR was possible only when POB-DNA was 5'-phosphorylated prior to the LMPCR procedure. NNKOAc generated damage at all four bases with the decreasing order guanine>adenine>cytosine>thymine. In contrast to NNKOAc damage distribution patterns, those induced by N-nitroso(acetoxymethyl)methylamine, a methylating NNK analog, induced damage principally at G positions detectable by enzymatic means that did not require phosphorylation. Analysis of damage distribution patterns, reveals a high frequency of damage in the p53 gene in codons 241 and 245 and a lower frequency of damage in codon 248. We analyzed the 3' termini of the NNKOAc induced single-strand breaks using a (32)P-post-labeling assay or a nucleotide exchange reaction at the 3'-termini catalyzed by T4 DNA polymerase combined with endonuclease IV treatment. Both methods indicate that the 3' termini of the single-strand breaks are not hydroxyl groups and are blocked by an unknown chemical structure that is not recognized by endonuclease IV. These data are consistent with POB-phosphotriester hydrolysis leading to strand breaks in DNA. The POB-damage could be mutagenic because NNKOAc produces single-strand breaks with the products being a 5'-hydroxyl group and a 3'-blocking group and strand breaks. These results represent the first step in determining if NNK pyridyloxobutylates DNA with sequence specificity similar to those observed with other model compounds.
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| pubmed:grant | |
| pubmed:language |
eng
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| pubmed:journal | |
| pubmed:citationSubset |
IM
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| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/4-(N-methyl-N-nitrosamino)-1-(3-pyri...,
http://linkedlifedata.com/resource/pubmed/chemical/Carcinogens,
http://linkedlifedata.com/resource/pubmed/chemical/DNA, Single-Stranded,
http://linkedlifedata.com/resource/pubmed/chemical/DNA Adducts,
http://linkedlifedata.com/resource/pubmed/chemical/Nitrosamines,
http://linkedlifedata.com/resource/pubmed/chemical/Nucleotides,
http://linkedlifedata.com/resource/pubmed/chemical/Phosphodiesterase I,
http://linkedlifedata.com/resource/pubmed/chemical/Phosphoric Diester Hydrolases
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| pubmed:status |
MEDLINE
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| pubmed:month |
Oct
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| pubmed:issn |
0022-2836
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| pubmed:author | |
| pubmed:copyrightInfo |
Copyright 2001 Academic Press.
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| pubmed:issnType |
Print
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| pubmed:day |
26
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| pubmed:volume |
313
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| pubmed:owner |
NLM
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| pubmed:authorsComplete |
Y
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| pubmed:pagination |
539-57
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| pubmed:dateRevised |
2007-11-14
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| pubmed:meshHeading |
pubmed-meshheading:11676538-Alkylation,
pubmed-meshheading:11676538-Base Sequence,
pubmed-meshheading:11676538-Carcinogens,
pubmed-meshheading:11676538-DNA, Single-Stranded,
pubmed-meshheading:11676538-DNA Adducts,
pubmed-meshheading:11676538-DNA Damage,
pubmed-meshheading:11676538-DNA Methylation,
pubmed-meshheading:11676538-DNA Mutational Analysis,
pubmed-meshheading:11676538-Escherichia coli,
pubmed-meshheading:11676538-Exons,
pubmed-meshheading:11676538-Genes, p53,
pubmed-meshheading:11676538-Genes, ras,
pubmed-meshheading:11676538-Genome, Human,
pubmed-meshheading:11676538-Humans,
pubmed-meshheading:11676538-Lung Neoplasms,
pubmed-meshheading:11676538-Molecular Sequence Data,
pubmed-meshheading:11676538-Mutagenesis,
pubmed-meshheading:11676538-Nitrosamines,
pubmed-meshheading:11676538-Nucleotides,
pubmed-meshheading:11676538-Phosphodiesterase I,
pubmed-meshheading:11676538-Phosphoric Diester Hydrolases,
pubmed-meshheading:11676538-Phosphorylation,
pubmed-meshheading:11676538-Polymerase Chain Reaction,
pubmed-meshheading:11676538-Smoking,
pubmed-meshheading:11676538-Substrate Specificity
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| pubmed:year |
2001
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| pubmed:articleTitle |
Characterization and mapping of DNA damage induced by reactive metabolites of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) at nucleotide resolution in human genomic DNA.
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| pubmed:affiliation |
Laboratory of Cancer Etiology and Chemoprevention, Faculty of Pharmacy, Laval University, Quebec City, G1K 7P4, Canada.
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| pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
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