Functionality of the dnaA protein binding site in DNA replication is orientation-dependent.
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.
Seufert
W
W
Max-Planck-Institut für Molekulare Genetik, Berlin, West Germany.
Dobrinski
B
B
Lurz
R
R
Messer
W
W
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
D001426:Bacterial Proteins; D004269:DNA, Bacterial