Role of high-fidelity Escherichia coli DNA polymerase I in replication bypass of a deoxyadenosine DNA-peptide cross-link

Kinrin Yamanaka, Irina G. Minko, Steven E. Finkel, Myron F. Goodman, R. Stephen Lloyd

Research output: Contribution to journalArticle

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Abstract

Reaction of bifunctional electrophiles with DNA in the presence of peptides can result in DNA-peptide cross-links. In particular, the linkage can be formed in the major groove of DNA via the exocyclic amino group of adenine (N 6-dA). We previously demonstrated that an A family human polymerase, Pol v, can efficiently and accurately synthesize DNA past N 6-dA-linked peptides. Based on these results, we hypothesized that another member of that family, Escherichia coli polymerase I (Pol I), may also be able to bypass these large major groove DNA lesions. To test this, oligodeoxynucleotides containing a site-specific N 6-dA dodecylpeptide crosslink were created and utilized for in vitro DNA replication assays using E. coli DNA polymerases. The results showed that Pol I and Pol II could efficiently and accurately bypass this adduct, while Pol III replicase, Pol IV, and Pol V were strongly inhibited. In addition, cellular studies were conducted using E. coli strains that were either wild type or deficient in all three DNA damage-inducible polymerases, i.e., Pol II, Pol IV, and Pol V. When single-stranded DNA vectors containing a site-specific N 6-dA dodecylpeptide cross-link were replicated in these strains, the efficiencies of replication were comparable, and in both strains, intracellular bypass of the lesion occurred in an error-free manner. Collectively, these findings demonstrate that despite its constrained active site, Pol I can catalyze DNA synthesis past N 6-dA-linked peptide cross-links and is likely to play an essential role in cellular bypass of large major groove DNA lesions.

Original languageEnglish (US)
Pages (from-to)3815-3821
Number of pages7
JournalJournal of bacteriology
Volume193
Issue number15
DOIs
StatePublished - Aug 1 2011

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ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

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