Open AccessApplication of DNA adductomics to soil bacterium Sphingobium sp. strain KK 22
Author(s) -
Kanaly Robert A.,
Micheletto Ruggero,
Matsuda Tomonari,
Utsuno Youko,
Ozeki Yasuhiro,
Hamamura Natsuko
Publication year - 2015
Publication title -
microbiologyopen
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.881
H-Index - 36
ISSN - 2045-8827
DOI - 10.1002/mbo3.283
Subject(s) - acrolein , dna , bacteria , strain (injury) , chemistry , adduct , dna damage , electrospray ionization , genotoxicity , mass spectrometry , tandem mass spectrometry , biochemistry , microbiology and biotechnology , chromatography , biology , toxicity , organic chemistry , genetics , anatomy , catalysis
Toward the development of ecotoxicology methods to investigate microbial markers of impacts of hydrocarbon processing activities, DNA adductomic analyses were conducted on a sphingomonad soil bacterium. From growing cells that were exposed or unexposed to acrolein, a commonly used biocide in hydraulic fracturing processes, DNA was extracted, digested to 2′‐deoxynucleosides and analyzed by liquid chromatography‐positive ionization electrospray‐tandem mass spectrometry in selected reaction monitoring mode transmitting the [M + H] + > [M + H − 116] + transition over 100 transitions. Overall data shown as DNA adductome maps revealed numerous putative DNA adducts under both conditions with some occurring specifically for each condition. Adductomic analyses of triplicate samples indicated that elevated levels of some targeted putative adducts occurred in exposed cells. Two exposure‐specific adducts were identified in exposed cells as 3‐(2′‐deoxyribosyl)‐5,6,7,8‐tetrahydro‐6‐hydroxy‐(and 8‐hydroxy‐)pyrimido[1,2‐ a ]‐ purine‐(3 H )‐one (6‐ and 8‐hydroxy‐PdG) following synthesis of authentic standards of these compounds and subsequent analyses. A time course experiment showed that 6‐ and 8‐hydroxy‐PdG were detected in bacterial DNA within 30 min of acrolein exposure but were not detected in unexposed cells. This work demonstrated the first application of DNA adductomics to examine DNA damage in a bacterium and sets a foundation for future work.