Requirement of Watson-Crick hydrogen bonding for DNA synthesis by yeast DNA polymerase η

M. Todd Washington, Sandra A. Helquist, Eric T. Kool, Louise Prakash, Satya Prakash

Research output: Contribution to journalArticle

78 Scopus citations

Abstract

Classical high-fidelity DNA polymerases discriminate between the correct and incorrect nucleotides by using geometric constraints imposed by the tight fit of the active site with the incipient base pair. Consequently, Watson-Crick (W-C) hydrogen bonding between the bases is not required for the efficiency and accuracy of DNA synthesis by these polymerases. DNA polymerase η (Polη) is a low-fidelity enzyme able to replicate through DNA lesions. Using difluorotoluene, a nonpolar isosteric analog of thymine unable to form W-C hydrogen bonds with adenine, we found that the efficiency and accuracy of nucleotide incorporation by Polη are severely impaired. From these observations, we suggest that W-C hydrogen bonding is required for DNA synthesis by Polη; in this regard, Polη differs strikingly from classical high-fidelity DNA polymerases.

Original languageEnglish (US)
Pages (from-to)5107-5112
Number of pages6
JournalMolecular and cellular biology
Volume23
Issue number14
DOIs
StatePublished - Jul 2003

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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