Dpo4 is hindered in extending a G·T mismatch by a reverse wobble

Jose Trincao, Robert E. Johnson, William T. Wolfle, Carlos R. Escalante, Satya Prakash, Louise Prakash, Aneel K. Aggarwal

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Abstract

The ability or inability of a DNA polymerase to extend a mispair directly affects the establishment of genomic mutations. We report here kinetic analyses of the ability of Dpo4, a Y-family polymerase from Sulfolobus solfataricus, to extend from all mispairs opposite a template G or T. Dpo4 is equally inefficient at extending these mispairs, which include, surprisingly, a G·T mispair expected to conform closely to Watson-Crick geometry. To elucidate the basis of this, we solved the structure of Dpo4 bound to G·T-mispaired primer template in the presence of an incoming nucleotide. As a control, we also determined the structure of Dpo4 bound to a matched A-T base pair at the primer terminus. The structures offer a basis for the low efficiency of Dpo4 in extending a G·T mispair: a reverse wobble that deflects the primer 3′-OH away from the incoming nucleotide.

Original languageEnglish (US)
Pages (from-to)457-462
Number of pages6
JournalNature Structural and Molecular Biology
Volume11
Issue number5
DOIs
StatePublished - May 1 2004

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ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

Cite this

Trincao, J., Johnson, R. E., Wolfle, W. T., Escalante, C. R., Prakash, S., Prakash, L., & Aggarwal, A. K. (2004). Dpo4 is hindered in extending a G·T mismatch by a reverse wobble. Nature Structural and Molecular Biology, 11(5), 457-462. https://doi.org/10.1038/nsmb755