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J. Biol. Chem., Vol. 263, Issue 6, 2719-2723, 02, 1988

Functionality of the dnaA protein binding site in DNA replication is orientation-dependent

W Seufert, B Dobrinski, R Lurz and W Messer
Max-Planck-Institut fur Molekulare Genetik, Berlin, West Germany.

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.
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