title: Assembly of a chromosomal replication machine: two DNA polymerases, a clamp loader, and sliding clamps in one holoenzyme particle. I. Organization of the clamp loader.
abstract: The gamma complex of DNA polymerase III holoenzyme, the replicase of Escherichia coli, couples ATP hydrolysis to the loading of beta sliding clamps onto primed DNA. The beta sliding clamp tethers the holoenzyme replicase to DNA for rapid and processive synthesis. In this report, the gamma complex has been constituted from its five different subunits. Size measurements and subunit stoichiometry studies show a composition of gamma 2 delta 1 delta' 1 1 chi 1 psi 1. Strong intersubunit contacts have been identified by gel filtration, and weaker contacts were identified by surface plasmon resonance measurements. An analogous tau complex has also been constituted and characterized; it is nearly as active as the gamma complex in clamp loading activity, but as shown in the fourth report of this series, it is at a disadvantage in binding the delta, delta', chi, and psi subunits when core is present (Xiao, H., Naktinis, V., and O'Donnell, M. (1995) J. Biol. Chem. 270, 13378-13383). The single copy subunits within the gamma complex provide the basis for the structural asymmetry inherent within DNA polymerase III holoenzyme.
authors.affiliation: Microbiology Department, Cornell University Medical College, New York, New York 10021, USA.
authors.forename: R
authors.initials: R
authors.lastname: Onrust
authors.forename: J
authors.initials: J
authors.lastname: Finkelstein
authors.forename: V
authors.initials: V
authors.lastname: Naktinis
authors.forename: J
authors.initials: J
authors.lastname: Turner
authors.forename: L
authors.initials: L
authors.lastname: Fang
authors.forename: M
authors.initials: M
authors.lastname: O'Donnell
mesh_terms: D002851:Chromatography, High Pressure Liquid; D002876:Chromosomes, Bacterial; D004258:DNA Polymerase III / Q000378:metabolism*; D004261:DNA Replication; D004269:DNA, Bacterial / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics
chemical_list: D004269:DNA, Bacterial; D004258:DNA Polymerase III