Abstract
A new group of error-prone DNA polymerases overcomes the blockage posed to normal DNA replication by damaged template bases, suggesting an active site with a loose, flexible pocket that accommodates aberrant DNA structures. We have determined a 2.8 Å resolution crystal structure of the Sulfolobus solfataricus Dbh protein, a DNA translesion polymerase closely related to Escherichia coli DNA polymerase IV and human polymerase κ. A high error rate is observed for the Dbh polymerase in a range of 10−2–10−3 for all 12 base substitution mispairs. The crystal structure of Dbh reveals an overall architecture resembling other DNA polymerases but has unique features that are likely to contribute to error-prone synthesis, including −1 frameshifting mutations.
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Acknowledgements
This work was supported by research grants from the U.S. Department of Energy (to T.E.) and the National Insitutes of Health (to M.F.G.). L.F.S and E.A.T. are supported by postdoctoral fellowships from the National Institutes of Health. We thank the staff of the Harvard/Armenise Structural Biology Center (Harvard Medical School) and beamlines X-12C and X-25 of the Macromolecular Crystallography Research Resource at the National Synchrotron Light Source (Upton, NY) for assistance with X-ray data collection. We thank R. Woodgate (NICDHD, NIH) for providing an expression clone for the Dbh protein. Crystallographic coordinates of a catalytic fragment of Dbh were kindly provided by T. Steitz, J. Pata and B.-L. Zhou (Yale University), and coordinates of S. cerevisiase Pol η were kindly provided by J. Tincao and A. Aggarwal (Mount Sinai School of Medicine).
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Silvian, L., Toth, E., Pham, P. et al. Crystal structure of a DinB family error-prone DNA polymerase from Sulfolobus solfataricus. Nat Struct Mol Biol 8, 984–989 (2001). https://doi.org/10.1038/nsb1101-984
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DOI: https://doi.org/10.1038/nsb1101-984
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