title: New structural snapshots provide molecular insights into the mechanism of high fidelity DNA synthesis.
abstract: Time-lapse X-ray crystallography allows visualization of intermediate structures during the DNA polymerase catalytic cycle. Employing time-lapse crystallography with human DNA polymerase β has recently allowed us to capture and solve novel intermediate structures that are not stable enough to be analyzed by traditional crystallography. The structures of these intermediates reveals exciting surprises about active site metal ions and enzyme conformational changes as the reaction proceeds from the ground state to product release. In this perspective, we provide an overview of recent advances in understanding the DNA polymerase nucleotidyl transferase reaction and highlight both the significance and mysteries of enzyme efficiency and specificity that remain to be solved.
authors.affiliation: Genome Integrity and Structural Biology Laboratory, National Institute of Environmental Health Sciences, National Institutes of Health, 111 T.W. Alexander Drive, Research Triangle Park, NC 27709, United States.
authors.forename: Bret D
authors.initials: BD
authors.lastname: Freudenthal
authors.affiliation: Genome Integrity and Structural Biology Laboratory, National Institute of Environmental Health Sciences, National Institutes of Health, 111 T.W. Alexander Drive, Research Triangle Park, NC 27709, United States.
authors.forename: William A
authors.initials: WA
authors.lastname: Beard
authors.affiliation: Genome Integrity and Structural Biology Laboratory, National Institute of Environmental Health Sciences, National Institutes of Health, 111 T.W. Alexander Drive, Research Triangle Park, NC 27709, United States. Electronic address: wilson5@niehs.nih.gov.
authors.forename: Samuel H
authors.initials: SH
authors.lastname: Wilson
mesh_terms: D020134:Catalytic Domain; D002413:Cations, Divalent; D018360:Crystallography, X-Ray / Q000295:instrumentation; D004247:DNA / Q000096:biosynthesis* / Q000737:chemistry; D019951:DNA Polymerase beta / Q000737:chemistry* / Q000378:metabolism; D004260:DNA Repair; D006801:Humans; D008274:Magnesium / Q000737:chemistry* / Q000378:metabolism; D008345:Manganese / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D009154:Mutation; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D013379:Substrate Specificity; D059008:Time-Lapse Imaging / Q000295:instrumentation
chemical_list: D002413:Cations, Divalent; D008345:Manganese; D004247:DNA; D019951:DNA Polymerase beta; D008274:Magnesium
keywords: Base excision repair; DNA polymerase; DNA repair; Fidelity; Function; Genome instability; Structure; X-family; X-ray crystallography