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Closed Access item Activation of Replication Origins in ϕ29-related Phages Requires the Recognition of Initiation Proteins to Specific Nucleoprotein Complexes

Authors:Freire, Raimundo
Serrano, Manuel
Salas, Margarita
Hermoso, José Miguel
Issue Date:29-Nov-1996
Publisher:American Society for Biochemistry and Molecular Biology
Citation:Journal of Biological Chemistry 271: 31000-31007 (1996)
Abstract:Protein p6 of Bacillus subtilis phage ϕ29 activates the initiation of viral DNA replication by forming a multimeric nucleoprotein complex at the origins of replication, located at both ends of the linear genome. This activation requires a precise positioning of the protein p6 array with respect to the initiation site. To investigate this activation mechanism, we have purified the ϕ29 protein p6 counterparts from the related phages Nf and GA-1 and analyzed the formation of complexes with DNA. In the homologous protein p6-DNA complexes the ϕ29 and Nf protein arrays showed an identical positioning, different than that of the GA-1 protein array. In contrast, in the heterologous complexes the protein showed a different arrangement except in the case of the Nf protein-ϕ29 DNA complex. We have also purified the proteins involved in the initiation of replication (terminal protein and DNA polymerase) from phages Nf and GA-1 and measured the ability of the different p6 proteins to activate homologous and heterologous replication origins. The results obtained indicate that the activation requires not only the formation of a specific nucleoprotein complex but also its specific recognition by the proteins involved in the initiation of DNA replication.
Publisher version (URL):http://dx.doi.org/10.1074/jbc.271.48.31000
URI:http://hdl.handle.net/10261/38816
ISSN:0021-9258
1083-351X
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