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Volume 270, Number 17, Issue of April 28, pp. 9792-9799, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.

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The Requirement for Molecular Chaperones in DNA Replication Is Reduced by the Mutation in P Gene, Which Weakens the Interaction between P Protein and DnaB Helicase

Igor Konieczny , Jaroslaw Marszalek

During the initiation of DNA replication, the host DnaB helicase is complexed with phage P protein in order to be properly positioned near the ori-O initiation complex. However, the P-DnaB interaction inhibits the activities of DnaB. Thus, the concerted action of bacterial heat shock proteins, DnaK, DnaJ, and GrpE, is required to activate the helicase. Wild-type phage cannot grow on the E. coli dnaB, dnaK, dnaJ, and grpE mutants. However, phage with a mutation in the P gene, is able to produce progeny in these mutants as well as in the wild-type bacteria. Purified mutant protein reveals a much lower affinity to DnaB than wild-type P, and the -DnaB complex is unstable. Also, a very low concentration of DnaK protein is sufficient to activate the helicase in a replication system based on dv dsDNA. In that system, the mutant DnaK756 protein, inactive in the P-dependent replication, revealed its activity in the -dependent reaction. The O-P-dependent replication system based on M13 ssDNA efficiently replicates DNA in the absence of any chaperone protein, unless P is substituted by the mutant protein. Data presented in this paper explain why phage is able to grow on wild-type and dnaK756 bacteria.

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