title: DNA polymerase activity at the single-molecule level.
abstract: DNA polymerases are essential enzymes responsible for replication and repair of DNA in all organisms. To replicate DNA with high fidelity, DNA polymerases must select the correct incoming nucleotide substrate during each cycle of nucleotide incorporation, in accordance with the templating base. When an incorrect nucleotide is sometimes inserted, the polymerase uses a separate 3'→5' exonuclease to remove the misincorporated base (proofreading). Large conformational rearrangements of the polymerase-DNA complex occur during both the nucleotide incorporation and proofreading steps. Single-molecule fluorescence spectroscopy provides a unique tool for observation of these dynamic conformational changes in real-time, without the need to synchronize a population of DNA-protein complexes.
authors.affiliation: Department of Molecular Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
authors.forename: Joshua P
authors.initials: JP
authors.lastname: Gill
authors.forename: Jun
authors.initials: J
authors.lastname: Wang
authors.forename: David P
authors.initials: DP
authors.lastname: Millar
mesh_terms: D000818:Animals; D004261:DNA Replication / Q000502:physiology; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry* / Q000378:metabolism* / Q000502:physiology*; D004789:Enzyme Activation; D006801:Humans; D008954:Models, Biological; D008958:Models, Molecular; D009711:Nucleotides / Q000378:metabolism; D011487:Protein Conformation
chemical_list: D009711:Nucleotides; D004259:DNA-Directed DNA Polymerase