Inhibition of vaccinia virus replication by peptide aptamers.
A20 protein is a major component of the vaccinia virus replication complex. It binds to the DNA polymerase E9, the uracil DNA glycosylase D4 and the primase/helicase D5, three proteins that are essential for viral DNA synthesis. The identification of molecules able to interact with the replication complex and inhibit its activity is a promising strategy for the design of new anti-orthopoxvirus drugs. In this study, we used a yeast genetic approach to select, from combinatorial libraries, 8-mers peptide aptamers that specifically interact with A20. From this screen, we isolated five peptide aptamers whose binding to A20 was confirmed by a glutathione S-transferase (GST) pull-down assay. Among those, we determined that peptide aptamer 72 binds to a central domain on A20. Interestingly, this region of A20 was previously shown to be important for its function in DNA replication. We next showed that vaccinia virus DNA synthesis was impaired in cells constitutively expressing peptide aptamer 72 and that virus production was inhibited in those cells. Thus, peptide aptamer 72 may be a useful tool for the development of new compounds specifically targeting poxvirus replication.
Institut de Recherches Biomédicales des Armées, CRSSA Emile Pardé, Grenoble, France.
Laurent
L
Saccucci
Jean-Marc
JM
Crance
Pierre
P
Colas
Marc
M
Bickle
Daniel
D
Garin
Frédéric
F
Iseni
D000998:Antiviral Agents / Q000494:pharmacology*; D052158:Aptamers, Peptide / Q000494:pharmacology*; D002460:Cell Line; D004279:DNA, Viral / Q000096:biosynthesis; D006801:Humans; D011485:Protein Binding; D025941:Protein Interaction Mapping; D020798:Two-Hybrid System Techniques; D014616:Vaccinia virus / Q000187:drug effects* / Q000502:physiology; D014764:Viral Proteins / Q000378:metabolism; D014779:Virus Replication / Q000187:drug effects*
C422803:A20R protein, vaccinia virus; D000998:Antiviral Agents; D052158:Aptamers, Peptide; D004279:DNA, Viral; D014764:Viral Proteins