Nucleic Acids Research Advance Access published online on October 2, 2007
Nucleic Acids Research, doi:10.1093/nar/gkm749
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Nucleic Acid Enzymes |
Involvement of phage 
29 DNA polymerase and terminal protein subdomains in conferring specificity during initiation of protein-primed DNA replication
Instituto de Biología Molecular ‘Eladio Viñuela’ (CSIC), Centro de Biología Molecular ‘Severo Ochoa’ (CSIC-UAM), Campus Universidad Autónoma, Canto Blanco, 28049 Madrid, Spain
*To whom correspondence should be addressed. Tel: +34 914978435; Fax: +34 914978490; Email: msalas{at}cbm.uam.es The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors.
Received July 31, 2007. Revised September 6, 2007. Accepted September 10, 2007.
To initiate 
29 DNA replication, the DNA polymerase has to form a complex with the homologous primer terminal protein (TP) that further recognizes the replication origins of the homologous TP-DNA placed at both ends of the linear genome. By means of chimerical proteins, constructed by swapping the priming domain of the related 29 and GA-1 TPs, we show that DNA polymerase can form catalytically active heterodimers exclusively with that chimerical TP containing the N-terminal part of the homologous TP, suggesting that the interaction between the polymerase TPR-1 subdomain and the TP N-terminal part is the one mainly responsible for the specificity between both proteins. We also show that the TP N-terminal part assists the proper binding of the priming domain at the polymerase active site. Additionally, a chimerical 29 DNA polymerase containing the GA-1 TPR-1 subdomain could use GA-1 TP, but only in the presence of 29 TP-DNA as template, indicating that parental TP recognition is mainly accomplished by the DNA polymerase. The sequential events occurring during initiation of bacteriophage protein-primed DNA replication are proposed.
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