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Int J Biol Sci 2009; 5(6):611-620. doi:10.7150/ijbs.5.611 This issue Cite

Research Paper

Nucleotide excision repair and recombination are engaged in repair of trans-4-hydroxy-2-nonenal adducts to DNA bases in Escherichia coli

Beata Janowska¹, Marek Komisarski¹, Paulina Prorok², Beata Sokołowska³, Jarosław Kuśmierek¹, Celina Janion¹, Barbara Tudek^{1,2 ✉}

1. Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5a, 02-106 Warsaw, Poland;
2. Institute of Genetics and Biotechnology, Warsaw University, Pawińskiego 5a, 02-106 Warsaw, Poland;
3. Medical Research Center, Polish Academy of Sciences, Pawinskiego 5, 02-106 Warsaw, Poland

✉ Correspondence to: Barbara Tudek, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5a, 02-106 Warsaw, Poland. Tel: (+4822) 592-3334; fax: (+4822) 592-2190; e-mail: tudekwaw.pl

Abstract

One of the major products of lipid peroxidation is trans-4-hydroxy-2-nonenal (HNE). HNE forms highly mutagenic and genotoxic adducts to all DNA bases. Using M13 phage lacZ system, we studied the mutagenesis and repair of HNE treated phage DNA in E. coli wild-type or uvrA, recA, and mutL mutants. These studies revealed that: (i) nucleotide excision and recombination, but not mismatch repair, are engaged in repair of HNE adducts when present in phage DNA replicating in E. coli strains; (ii) in the single uvrA mutant, phage survival was drastically decreased while mutation frequency increased, and recombination events constituted 48 % of all mutations; (iii) in the single recA mutant, the survival and mutation frequency of HNE-modified M13 phage was slightly elevated in comparison to that in the wild-type bacteria. The majority of mutations in recA^- strain were G:C → T:A transversions, occurring within the sequence which in recA⁺ strains underwent RecA-mediated recombination, and the entire sequence was deleted; (iv) in the double uvrA recA mutant, phage survival was the same as in the wild-type although the mutation frequency was higher than in the wild-type and recA single mutant, but lower than in the single uvrA mutant. The majority of mutations found in the latter strain were base substitutions, with G:C → A:T transitions prevailing. These transitions could have resulted from high reactivity of HNE with G and C, and induction of SOS-independent mutations.

Keywords: trans-4-hydroxy-2-nonenal, HNE-DNA adducts, mutations, recombination, NER, M13 phage

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