title: The tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) induces mitochondrial and nuclear DNA damage in Caenorhabditis elegans.
abstract: The metabolites of the tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) form DNA adducts in animal models. While there are many reports of formation of nuclear DNA adducts, one report also detected NNK-induced damage to the mitochondrial genome in rats. Using a different DNA damage detection technology, we tested whether this finding could be repeated in the nematode Caenorhabditis elegans. We treated N2 strain (wild-type) nematodes with NNK in liquid culture, and applied quantitative PCR to analyze NNK-induced nuclear and mitochondrial DNA (mtDNA) damage. Our results confirm that NNK causes both nuclear and mtDNA damage. However, we did not detect a difference in the level of nuclear versus mtDNA damage in C. elegans. To test whether the mtDNA damage was associated with mitochondrial dysfunction, we used a transgenic nematode strain that permits in vivo measurement of ATP levels and found lower levels of ATP in NNK-exposed animals when compared with the unexposed controls. To test whether the lower levels of ATP could be attributed to inhibition of respiratory chain components, we investigated oxygen consumption in whole C. elegans and found reduced oxygen consumption in exposed animals when compared with the unexposed controls. Our data suggest a model in which NNK exposure causes damage to both C. elegans nuclear and mitochondrial genomes, and support the hypothesis that the mitochondrial damage is functionally important in this model. These results also represent a first step in developing this genetically tractable organism as a model for assessing NNK toxicity.
authors.affiliation: Nicholas School of the Environment, Duke University, Durham, North Carolina.
authors.forename: Rakesh
authors.initials: R
authors.lastname: Bodhicharla
authors.forename: Ian T
authors.initials: IT
authors.lastname: Ryde
authors.forename: G L
authors.initials: GL
authors.lastname: Prasad
authors.forename: Joel N
authors.initials: JN
authors.lastname: Meyer
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D017173:Caenorhabditis elegans / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development; D002467:Cell Nucleus / Q000187:drug effects* / Q000235:genetics; D004249:DNA Damage / Q000187:drug effects*; D004272:DNA, Mitochondrial; D008928:Mitochondria / Q000187:drug effects* / Q000235:genetics; D009602:Nitrosamines / Q000633:toxicity*; D010100:Oxygen / Q000378:metabolism; D014026:Nicotiana
chemical_list: D004272:DNA, Mitochondrial; D009602:Nitrosamines; C016583:4-(N-methyl-N-nitrosamino)-1-(3-pyridyl)-1-butanone; D000255:Adenosine Triphosphate; D010100:Oxygen
keywords: Caenorhabditis elegans; NNK; mitochondrial and nuclear DNA damage; tobacco