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Please use this identifier to cite or link to this item: http://hdl.handle.net/10261/37712
Title: A specific subdomain in 29 DNA polymerase confers both processivity and strand displacement capacity
Authors: Rodríguez García, Irene; Lázaro, José M.; Blanco, Luis; Kamtekar, S.; Berman, Andrea J.; Wang, J.; Steitz, T. A.; Salas, Margarita; Vega, Miguel de
Keywords: Protein-primed replication
Terminal protein region
Helicase-like activity
DNA-binding stability
Issue Date: 3-May-2005
Publisher: National Academy of Sciences (U.S.)
Citation: Proceedings of the National Academy of Sciences 102: 6407-6412 (2005)
Abstract: Recent crystallographic studies of φ29 DNA polymerase have provided structural insights into its strand displacement and processivity. A specific insertion named terminal protein region 2 (TPR2), present only in protein-primed DNA polymerases, together with the exonuclease, thumb, and palm subdomains, forms two tori capable of interacting with DNA. To analyze the functional role of this insertion, we constructed a φ29 DNA polymerase deletion mutant lacking TPR2 amino acid residues Asp-398 to Glu-420. Biochemical analysis of the mutant DNA polymerase indicates that its DNA-binding capacity is diminished, drastically decreasing its processivity. In addition, removal of the TPR2 insertion abolishes the intrinsic capacity of φ29 DNA polymerase to perform strand displacement coupled to DNA synthesis. Therefore, the biochemical results described here directly demonstrate that TPR2 plays a critical role in strand displacement and processivity.
Publisher version (URL): http://dx.doi.org/10.1073/pnas.0500597102
URI: http://hdl.handle.net/10261/37712
ISSN: 0027-8424
DOI: 10.1073/pnas.0500597102
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