Transcriptional activation of the origin of coliphage lambda DNA replication is regulated by the host DnaA initiator function.
The initiator of phage lambda DNA replication, the lambda O protein, is considered to be an analogue of the initiator of DNA replication (DnaA) of its host, Escherichia coli. Both specifically recognize their origins of replication, ori lambda and oriC, respectively, and organize the assembly of specific replication complexes. However, DnaA has an additional activation function, acting on oriC-proximal DnaA-boxes, and regulating transcription initiated at promoters in and around oriC. Here, we demonstrate that lambda plasmid replication can be synchronized by a temperature shift-down that caused renaturation of the previously denatured DnaAts protein. Moreover, we show that elimination of the activating DnaA function affects transcriptional activation at ori lambda. DnaA may act by binding to DnaA-boxes, situated around the lambda pR promoter; there are no such sequences in ori lambda. Our results being to explain in molecular terms why lambda plasmid replication is DnaA-dependent [Kur et al., J. Mol. Biol. 198 (1987) 203-210] and why the initiation of phage lambda DNA replication is blocked (in E. coli devoid of prophage Rac) after inactivation of DnaA [Wegrzyn et al., Genetics (1995) in press].
Wegrzyn
G
G
Department of Molecular Biology, University of Gdańsk, Poland.
Szalewska-Pałasz
A
A
Wegrzyn
A
A
Obuchowski
M
M
Taylor
K
K
D001426:Bacterial Proteins / Q000502:physiology*; D010582:Bacteriophage lambda / Q000235:genetics* / Q000502:physiology; D004261:DNA Replication; D004268:DNA-Binding Proteins / Q000502:physiology*; D004926:Escherichia coli / Q000378:metabolism / Q000821:virology*; D015964:Gene Expression Regulation, Bacterial; D012093:Replicon; D013696:Temperature; D014158:Transcription, Genetic; D014779:Virus Replication
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; C066312:DnaA protein, Bacteria