The interaction of bacteriophage P2 B protein with Escherichia coli DnaB helicase.
Bacteriophage P2 requires several host proteins for lytic replication, including helicase DnaB but not the helicase loader, DnaC. Some genetic studies have suggested that the loading is done by a phage-encoded protein, P2 B. However, a P2 minichromosome containing only the P2 initiator gene A and a marker gene can be established as a plasmid without requiring the P2 B gene. Here we demonstrate that P2 B associates with DnaB. This was done by using the yeast two-hybrid system in vivo and was confirmed in vitro, where (35)S-labeled P2 B bound specifically to DnaB adsorbed to Q Sepharose beads and monoclonal antibodies directed against the His-tagged P2 B protein were shown to coprecipitate the DnaB protein. Finally, P2 B was shown to stabilize the opening of a reporter origin, a reaction that is facilitated by the inactivation of DnaB. In this respect, P2 B was comparable to lambda P protein, which is known to be capable of binding and inactivating the helicase while acting as a helicase loader. Even though P2 B has little similarity to other known or predicted helicase loaders, we suggest that P2 B is required for efficient loading of DnaB and that this role, although dispensable for P2 plasmid replication, becomes essential for P2 lytic replication.
Odegrip
R
R
Department of Genetics, Stockholm University, S-10691 Stockholm, Sweden.
Schoen
S
S
Haggård-Ljungquist
E
E
Park
K
K
Chattoraj
D K
DK
D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D017901:Bacteriophage P2 / Q000378:metabolism*; D003001:Cloning, Molecular; D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism*; D053488:DnaB Helicases; D004926:Escherichia coli / Q000201:enzymology*; D008969:Molecular Sequence Data; D010957:Plasmids; D012441:Saccharomyces cerevisiae; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
C407903:B protein, Enterobacteria phage P2; D001426:Bacterial Proteins; D014764:Viral Proteins; D004265:DNA Helicases; D053488:DnaB Helicases