Oxidation of 7,8-dihydro-8-oxoguanine affords lesions that are potent sources of replication errors in vivo

Paul Henderson, James C. Delaney, Feng Gu, Steven R. Tannenbaum, John M. Essigmann

Research output: Contribution to journalArticle

121 Scopus citations

Abstract

Three single-stranded DNA genomes have been constructed that contain the 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodG) oxidation products oxaluric acid, oxazalone, and cyanuric acid. Oligonucleotides containing each lesion were synthesized by treating an oligonucleotide containing a single 8-oxodG with peroxynitrite, and the desired products were isolated by HPLC. The modified oligonucleotides were ligated into M13mp7L2 bacteriophage DNA in such a way that the lesion was situated at a known site in the lacZ gene fragment of the viral genome. The circular genomes were transfected into wild-type AB1157 Escherichia coli. The relative efficiency of lesion bypass by DNA polymerase was determined by counting the number of initial independent infections produced by each genome relative to that of an unmodified DNA control. Viral progeny were analyzed for mutation frequency and type by PCR amplification of the insert region followed by a recently developed post-labeling assay. All three secondary lesions were readily bypassed, causing G → T transversions at frequencies at least an order of magnitude higher than 8-oxodG. These data establish a model whereby the modestly mutagenic primary lesion 8-oxodG is oxidized in vivo to much more highly mutagenic secondary lesions.

Original languageEnglish (US)
Pages (from-to)914-921
Number of pages8
JournalBiochemistry
Volume41
Issue number3
DOIs
StatePublished - Jan 22 2002
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry

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