Please use this identifier to cite or link to this item: http://hdl.handle.net/10261/37794
Share/Impact:
Título : Phage φ29 protein p56 prevents viral DNA replication impairment caused by uracil excision activity of uracil-DNA glycosylase
Autor : Serrano-Heras, Gemma, Bravo García, Alicia, Salas, Margarita
Palabras clave : φ29 DNA polymerase
DNA repair
dUMP incorporation
Protein-primed replication
Fecha de publicación : 9-Oct-2008
Editor: National Academy of Sciences (U.S.)
Citación : Proceedings of the National Academy of Sciences 105(2): 19044-19049 (2008)
Resumen: Protein p56 encoded by the Bacillus subtilis phage φ29 inhibits host uracil-DNA glycosylase (UDG) activity. In previous studies, we suggested that this inhibition is likely a defense mechanism developed by phage φ29 to prevent the action of UDG if uracilation occurs in DNA either from deamination of cytosine or the incorporation of dUMP during viral DNA replication. In this work, we analyzed the ability of φ29 DNA polymerase to insert dUMP into DNA. Primer extension analysis showed that viral DNA polymerase incorporates dU opposite dA with a catalytic efficiency only 2-fold lower than that for dT. Using the φ29 DNA amplification system, we found that φ29 DNA polymerase is also able to carry out the extension of the dA:dUMP pair and replicate past uracil. Additionally, UDG and apurinic-apyrimidinic endonuclease treatment of viral DNA isolated from φ29-infected cells revealed that uracil residues arise in φ29 DNA during replication, probably as a result of misincorporation of dUMP by the φ29 DNA polymerase. On the other hand, the action of UDG on uracil-containing φ29 DNA impaired in vitro viral DNA replication, which was prevented by the presence of protein p56. Furthermore, transfection activity of uracil-containing φ29 DNA was significantly higher in cells that constitutively synthesized p56 than in cells lacking this protein. Thus, our data support a model in which protein p56 ensures an efficient viral DNA replication, preventing the deleterious effect caused by UDG when it eliminates uracil residues present in the φ29 genome.
Versión del editor: http://dx.doi.org/10.1073/pnas.0808797105
URI : http://hdl.handle.net/10261/37794
DOI: 10.1073/pnas.0808797105
Citación : Proceedings of the National Academy of Sciences 105(2): 19044-19049 (2008)
Appears in Collections:(CBM) Artículos
(CIB) Artículos

Files in This Item:
File Description SizeFormat 
PNAS-2008-Serrano-Heras-19044-9.pdf417,15 kBAdobe PDFView/Open
Show full item record
 
CSIC SFX LinksSFX Query


Items in Digital.CSIC are protected by copyright, with all rights reserved, unless otherwise indicated.