Abstract
Innovative advances in X-ray crystallography and single-molecule biophysics have yielded unprecedented insight into the mechanisms of DNA lesion bypass and damage repair. Time-dependent X-ray crystallography has been successfully applied to view the bypass of 8-oxo-7,8-dihydro-2'-deoxyguanine (8-oxoG), a major oxidative DNA lesion, and the incorporation of the triphosphate form, 8-oxo-dGTP, catalyzed by human DNA polymerase β. Significant findings of these studies are highlighted here, and their contributions to the current mechanistic understanding of mutagenic translesion DNA synthesis (TLS) and base excision repair are discussed. In addition, single-molecule Forster resonance energy transfer (smFRET) techniques have recently been adapted to investigate nucleotide binding and incorporation opposite undamaged dG and 8-oxoG by Sulfolobus solfataricus DNA polymerase IV (Dpo4), a model Y-family DNA polymerase. The mechanistic response of Dpo4 to a DNA lesion and the complex smFRET technique are described here. In this perspective, we also describe how time-dependent X-ray crystallography and smFRET can be used to achieve the spatial and temporal resolutions necessary to answer some of the mechanistic questions that remain in the fields of TLS and DNA damage repair.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 260-269 |
| Number of pages | 10 |
| Journal | Chemical research in toxicology |
| Volume | 30 |
| Issue number | 1 |
| DOIs | |
| State | Published - Jan 17 2017 |
| Externally published | Yes |
Bibliographical note
Funding Information:Corresponding Author *Tel: +1 614 6883706. Fax: +1 614 2926773. E-mail: suo.3@osu. edu. Author Contributions †A.T.R. and A.J.R. contributed equally to this work. Funding This work was supported by the National Institutes of Health (Grants ES024585 and ES026821 to Z.S. and T32GM008512 to A.T.R.) and the National Science Foundation (Grant MCB-0960961 to Z.S.). Notes The authors declare no competing financial interest. Biographies Austin T. Raper graduated Summa Cum Laude from the University of Mount Union in 2013 with a B.S. in Biochemistry. As a fellow of the Chemistry and Biology Interface Training Program and member of the Ohio State Biochemistry Program, he is currently conducting research in the laboratory of Dr. Zucai Suo (Department of Chemistry and Biochemistry) at The Ohio State University to earn a Ph.D. in Biochemistry. Austin’s research focuses on characterizing enzymes critical for DNA lesion bypass and damage repair using biophysical and presteady-state kinetic methodologies.
Funding Information:
This work was supported by the National Institutes of Health (Grants ES024585 and ES026821 to Z.S. and T32GM008512 to A.T.R.) and the National Science Foundation (Grant MCB-0960961 to Z.S.).
Publisher Copyright:
© 2016 American Chemical Society.