title: Functionality of the dnaA protein binding site in DNA replication is orientation-dependent.
abstract: We analyzed the functionality of different dnaA protein binding sites by assaying in vitro dnaA-dependent replication of pBR322 derivatives. Single dnaA sites from oriC and from the mioC and dnaA gene promoters were active when combined with the primer generating element of pBR322 in a proper distance. Prereplisome assembly did not require sequences in addition to the 9-base pair consensus dnaA binding site. Inversion of the structurally asymmetric dnaA site relative to its orientation in wild type pBR322 resulted in a marked reduction in replication efficiency, as observed with five different dnaA sites studied. The direction of DNA replication was not affected.
authors.affiliation: Max-Planck-Institut für Molekulare Genetik, Berlin, West Germany.
authors.forename: W
authors.initials: W
authors.lastname: Seufert
authors.forename: B
authors.initials: B
authors.lastname: Dobrinski
authors.forename: R
authors.initials: R
authors.lastname: Lurz
authors.forename: W
authors.initials: W
authors.lastname: Messer
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism*; D001665:Binding Sites; D004261:DNA Replication; D004269:DNA, Bacterial / Q000096:biosynthesis*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011487:Protein Conformation
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial