title: Structure of PolC reveals unique DNA binding and fidelity determinants.
abstract: PolC is the polymerase responsible for genome duplication in many Gram-positive bacteria and represents an attractive target for antibacterial development. We have determined the 2.4-A resolution crystal structure of Geobacillus kaustophilus PolC in a ternary complex with DNA and dGTP. The structure reveals nascent base pair interactions that lead to highly accurate nucleotide incorporation. A unique beta-strand motif in the PolC thumb domain contacts the minor groove, allowing replication errors to be sensed up to 8 nt upstream of the active site. PolC exhibits the potential for large-scale conformational flexibility, which could encompass the catalytic residues. The structure suggests a mechanism by which the active site can communicate with the rest of the replisome to trigger proofreading after nucleotide misincorporation, leading to an integrated model for controlling the dynamic switch between replicative and repair polymerases. This ternary complex of a cellular replicative polymerase affords insights into polymerase fidelity, evolution, and structural diversity.
authors.affiliation: Replidyne, Inc., Louisville, CO 80027, USA.
authors.forename: Ronald J
authors.initials: RJ
authors.lastname: Evans
authors.forename: Douglas R
authors.initials: DR
authors.lastname: Davies
authors.forename: James M
authors.initials: JM
authors.lastname: Bullard
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Christensen
authors.forename: Louis S
authors.initials: LS
authors.lastname: Green
authors.forename: Joseph W
authors.initials: JW
authors.lastname: Guiles
authors.forename: Janice D
authors.initials: JD
authors.lastname: Pata
authors.forename: Wendy K
authors.initials: WK
authors.lastname: Ribble
authors.forename: Nebojsa
authors.initials: N
authors.lastname: Janjic
authors.forename: Thale C
authors.initials: TC
authors.lastname: Jarvis
mesh_terms: D020816:Amino Acid Motifs / Q000502:physiology; D001406:Bacillaceae / Q000201:enzymology*; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D020134:Catalytic Domain / Q000502:physiology; D018360:Crystallography, X-Ray; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry* / Q000378:metabolism; D016680:Genome, Bacterial / Q000502:physiology; D020836:Protein Structure, Quaternary / Q000502:physiology; D017434:Protein Structure, Tertiary / Q000502:physiology; D013329:Structure-Activity Relationship
chemical_list: D001426:Bacterial Proteins; C446929:PolC protein, bacteria; D004259:DNA-Directed DNA Polymerase