Involvement of phage φ29 DNA polymerase and terminal protein subdomains in conferring specificity during initiation of protein-primed DNA replication
Entity
UAM. Departamento de Biología MolecularPublisher
Oxford University Press [University Publisher]Date
2007-10-02Citation
10.1093/nar/gkm749
Nucleic Acids Research 35.21 (2007): 7061-7073
ISSN
0305-1048DOI
10.1093/nar/gkm749Project
Gobierno de España. BFU 2005-00733Editor's Version
10.1093/nar/gkm749Subjects
DNA Directed DNA Polymerase; Bacteriophages; Exonucleases; Biología y Biomedicina / BiologíaNote
This is an electronic version of an article published in Nucleic Acids Research. Patricia Pérez-Arnaiz, Elisa Longás,Laurentino Villar, José M. Lázaro, Margarita Salas and Miguel de Vega. Involvement of phage φ29 DNA polymerase and terminal protein subdomains in conferring specificity during initiation of protein-primed DNA replication. Nucleic Acids Research 35.21 (2007): 7061-7073Rights
2007 The Author(s)Abstract
To initiate φ29 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 φ29 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 φ29 DNA polymerase containing the GA-1 TPR-1 subdomain could use GA-1 TP, but only in the presence of φ29 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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Google Scholar:Pérez Arnáiz, Patricia
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Longás Torné, Elisa
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Villar, Laurentino
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Lázaro, José M.
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Salas, Margarita
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Vega, Miguel de
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