Accuracy of lesion bypass by yeast and human DNA polymerase η

M. Todd Washington, Robert E. Johnson, Louise Prakash, Satya Prakash

Research output: Contribution to journalArticle

62 Citations (Scopus)

Abstract

DNA polymerase η (Polη) functions in the error-free bypass of UV-induced DNA lesions, and a defect in Polη in humans causes the cancer-prone syndrome, the variant form of xeroderma pigmentosum. Both yeast and human Polη replicate through a cis-syn thymine-thymine dimer (TT dimer) by inserting two As opposite the two Ts of the dimer. Polη however, is a low-fidelity enzyme, and it misinserts nucleotides with a frequency of ≈ 10-2 to 10-3 opposite the two Ts of the TT dimer as well as opposite the undamaged template bases. This low fidelity of nucleotide insertion seems to conflict with the role of Polη in the error-free bypass of UV lesions. To resolve this issue, we have examined the ability of human and yeast Polη to extend from paired and mispaired primer termini opposite a TT dimer by using steady-state kinetic assays. We find that Polη extends from mispaired primer termini on damaged and undamaged DNAs with a frequency of ≈ 10-2 to 10-3 relative to paired primer termini. Thus, after the incorporation of an incorrect nucleotide, Polη would dissociate from the DNA rather than extend from the mispair. The resulting primer-terminal mispair then could be subject to proofreading by a 3′→5′ exonuclease. Replication through a TT dimer by Polη then would be more accurate than that predicted from the fidelity of nucleotide incorporation alone.

Original languageEnglish (US)
Pages (from-to)8355-8360
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume98
Issue number15
DOIs
StatePublished - Jul 17 2001

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Pyrimidine Dimers
Thymine
DNA-Directed DNA Polymerase
Nucleotides
Yeasts
DNA
Exonucleases
Enzymes
Neoplasms

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Accuracy of lesion bypass by yeast and human DNA polymerase η. / Washington, M. Todd; Johnson, Robert E.; Prakash, Louise; Prakash, Satya.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 98, No. 15, 17.07.2001, p. 8355-8360.

Research output: Contribution to journalArticle

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