Nucleic Acids Research Advance Access published online on August 13, 2007
Nucleic Acids Research, doi:10.1093/nar/gkm584
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Molecular Biology |
Protein p56 from the Bacillus subtilis phage
29 inhibits DNA-binding ability of uracil-DNA glycosylase
1Instituto de Biología Molecular Eladio Viñuela (CSIC), Centro de Biología Molecular Severo Ochoa (CSIC-UAM), Universidad Autónoma, Cantoblanco, 28049 Madrid and 2Centro de Investigaciones Biológicas (CSIC), Ramiro de Maeztu 9, 28040 Madrid, Spain
*To whom correspondence should be addressed. Tel: +34 91 497 8435; Fax: +34 91 497 8490; Email: msalas{at}cbm.uam.es
Received April 27, 2007. Revised July 16, 2007. Accepted July 17, 2007.
Protein p56 (56 amino acids) from the Bacillus subtilis phage
29 inactivates the host uracil-DNA glycosylase (UDG), an enzyme involved in the base excision repair pathway. At present, p56 is the only known example of a UDG inhibitor encoded by a non-uracil containing viral DNA. Using analytical ultracentrifugation methods, we found that protein p56 formed dimers at physiological concentrations. In addition, circular dichroism spectroscopic analyses revealed that protein p56 had a high content of ß-strands (around 40%). To understand the mechanism underlying UDG inhibition by p56, we carried out in vitro experiments using the Escherichia coli UDG enzyme. The highly acidic protein p56 was able to compete with DNA for binding to UDG. Moreover, the interaction between p56 and UDG blocked DNA binding by UDG. We also demonstrated that Ugi, a protein that interacts with the DNA-binding domain of UDG, was able to replace protein p56 previously bound to the UDG enzyme. These results suggest that protein p56 could be a novel naturally occurring DNA mimicry.
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G. Serrano-Heras, A. Bravo, and M. Salas Inaugural Article: Phage {varphi}29 protein p56 prevents viral DNA replication impairment caused by uracil excision activity of uracil-DNA glycosylase PNAS, December 9, 2008; 105(49): 19044 - 19049. [Abstract] [Full Text] [PDF] |
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