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. 1994 Dec 11;22(24):5204–5210. doi: 10.1093/nar/22.24.5204

Studies of bacteriophage P2 DNA replication: localization of the cleavage site of the A protein.

Y Liu 1, E Haggård-Ljungquist 1
PMCID: PMC332061  PMID: 7816607

Abstract

Bacteriophage P2 replicates via a modified rolling circle-type of mechanism, where the P2 A protein acts as an initiator of the replication by inducing a single-stranded cut at the origin of replication (ori). The exact location of the cut induced by the A protein in vivo is determined in this report by: (i) restriction analysis; (ii) DNA sequence analysis; and (iii) primer extensions. It is located 89.2% from the left end of the P2 genome, which is within the coding part of the A gene, in a region devoid of secondary structures. The A gene has been cloned into an expression vector, and the A protein has been purified. The purified A protein does not bind to double-stranded ori containing DNA, but it cleaves single-stranded ori containing DNA, which indicates that a special DNA structure and/or protein is required to make the ori accessible for the A protein.

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Selected References

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