Nucleic Acids Research Advance Access published online on October 28, 2006
Nucleic Acids Research, doi:10.1093/nar/gkl769
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© 2006 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Nucleic Acid Enzymes |
Functional characterization of highly processive protein-primed DNA polymerases from phages Nf and GA-1, endowed with a potent strand displacement capacity
Instituto de Biología Molecular ‘Eladio Viñuela’ (CSIC), Centro de Biología Molecular ‘Severo Ochoa’ (CSIC-UAM) Universidad Autónoma, Canto Blanco, 28049 Madrid, Spain
*To whom correspondence should be addressed. Tel: +344 91 4978436; Fax: +34 91 4978490; Email: msalas{at}cbm.uam.es
Received July 21, 2006. Revised September 11, 2006. Accepted September 28, 2006.
This paper shows that the protein-primed DNA polymerases encoded by bacteriophages Nf and GA-1, unlike other DNA polymerases, do not require unwinding or processivity factors for efficient synthesis of full-length terminal protein (TP)-DNA. Analysis of their polymerization activity shows that both DNA polymerases base their replication efficiency on a high processivity and on the capacity to couple polymerization to strand displacement. Both enzymes are endowed with a proofreading activity that acts coordinately with the polymerization one to edit polymerization errors. Additionally, Nf double-stranded DNA binding protein (DBP) greatly stimulated the in vitro formation of the TP-dAMP initiation complex by decreasing the Km value for dATP of the Nf DNA polymerase by >20-fold. Whereas Nf DNA polymerase, as the 
29 enzyme, is able to use its homologous TP as well as DNA as primer, GA-1 DNA polymerase appears to have evolved to use its corresponding TP as the only primer of DNA synthesis. Such exceptional behaviour is discussed in the light of the recently solved structure of the DNA polymerase/TP complex of the related bacteriophage 29.
The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors
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