PMID:10756017
| Citation |
Odegrip, R, Schoen, S, Haggård-Ljungquist, E, Park, K and Chattoraj, DK (2000) The interaction of bacteriophage P2 B protein with Escherichia coli DnaB helicase. J. Virol. 74:4057-63 |
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| Abstract |
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. |
| Links | |
| Keywords |
Amino Acid Sequence; Bacterial Proteins; Bacteriophage P2/metabolism; Cloning, Molecular; DNA Helicases/genetics; DNA Helicases/metabolism; DnaB Helicases; Escherichia coli/enzymology; Molecular Sequence Data; Plasmids; Saccharomyces cerevisiae; Viral Proteins/genetics; Viral Proteins/metabolism |
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