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Open Access item A specific subdomain in 29 DNA polymerase confers both processivity and strand displacement capacity
|Authors:||Rodríguez García, Irene|
Lázaro, José M.
Steitz, T. A.
Vega, Miguel de
|Keywords:||Protein-primed replication, Terminal protein region, Helicase-like activity, DNA-binding stability|
|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|
|Appears in Collections:||(CBM) Artículos|
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