Switch from theta to sigma replication of bacteriophage lambda DNA: factors involved in the process and a model for its regulation.
Bacteriophage lambda genome is one of the classical model replicons in studies on the regulation of DNA replication. Moreover, since genes coding for Shiga toxins are located in genomes of lambdoid phages, understanding of mechanisms controlling lambda DNA replication may be of bio-medical importance. During lytic development of bacteriophage lambda, its genome is replicated according to the theta (circle-to-circle) mode early after infection, and then it is switched to the sigma (rolling circle) mode. Two mechanisms of regulation of this switch were proposed recently and both suggested a crucial role for directionality of lambda DNA replication. Whereas one hypothesis assumed transient impairment of ClpP/ClpX-mediated proteolysis of the lambdaO initiator protein, another suggested a crucial role for transcriptional activation of the orilambda region and factors involved in the control of the p (R) promoter activity. Here we demonstrate that mutations in clpP and clpX genes had little influence on both directionality of lambda DNA replication and appearance of sigma replication intermediates. On the other hand, regulators affecting activity of the p (R) promoter (responsible for initiation of transcription, which activates orilambda) directly or indirectly influenced directionality of lambda DNA replication to various extents. Therefore, we conclude that regulation of the efficiency of transcriptional activation of orilambda, rather than transient impairment of the lambdaO proteolysis, is responsible for the control of the switch from theta to sigma replication, and propose a model for this control.
Narajczyk
Magdalena
M
Department of Molecular Biology, University of Gdańsk, Kładki 24, 80-822, Gdańsk, Poland.
Barańska
Sylwia
S
Wegrzyn
Alicja
A
Wegrzyn
Grzegorz
G
D000074183:ATPases Associated with Diverse Cellular Activities; D000251:Adenosine Triphosphatases / Q000235:genetics; D010582:Bacteriophage lambda / Q000235:genetics*; D004261:DNA Replication; D004279:DNA, Viral / Q000235:genetics*; D004587:Electrophoresis, Agar Gel; D049071:Endopeptidase Clp / Q000235:genetics; D004926:Escherichia coli / Q000378:metabolism / Q000821:virology; D029968:Escherichia coli Proteins / Q000235:genetics; D015967:Gene Expression Regulation, Viral; D008957:Models, Genetic; D018832:Molecular Chaperones / Q000235:genetics; D009154:Mutation; D010957:Plasmids / Q000378:metabolism; D018741:Replication Origin; D014158:Transcription, Genetic; D015533:Transcriptional Activation
D004279:DNA, Viral; D029968:Escherichia coli Proteins; D018832:Molecular Chaperones; C483572:ClpP protease, E coli; D049071:Endopeptidase Clp; D000251:Adenosine Triphosphatases; C088072:ClpX protein, E coli; D000074183:ATPases Associated with Diverse Cellular Activities