1. Inhibition of vaccinia virus replication by peptide aptamers
- Author
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Daniel Garin, Laurent Saccucci, Pierre Colas, Marc Bickle, Frédéric Iseni, Jean-Marc Crance, Centre international d'études supérieures en sciences agronomiques (Montpellier SupAgro), Institut de Recherches sur les lois Fondamentales de l'Univers (IRFU), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay, Aptanomics, Laboratoire de virologie du Centre de Recherches du Service de Santé des Armées (CRSSA), Ministère de la Défense, Virologie et pathogenèse virale (VPV), Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon, Université Claude Bernard Lyon 1 (UCBL), and Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)
- Subjects
MESH: Antiviral Agents ,Aptamer ,Vaccinia virus ,Eukaryotic DNA replication ,[SDV.BC]Life Sciences [q-bio]/Cellular Biology ,Biology ,Virus Replication ,MESH: Two-Hybrid System Techniques ,Antiviral Agents ,Cell Line ,DNA replication factor CDT1 ,Viral Proteins ,03 medical and health sciences ,Replication factor C ,Two-Hybrid System Techniques ,Virology ,hemic and lymphatic diseases ,Protein Interaction Mapping ,MESH: Vaccinia virus ,Humans ,MESH: Protein Binding ,Replication protein A ,030304 developmental biology ,Pharmacology ,0303 health sciences ,MESH: Humans ,030306 microbiology ,MESH: Virus Replication ,MESH: Protein Interaction Mapping ,DNA replication ,MESH: Aptamers, Peptide ,Molecular biology ,MESH: Viral Proteins ,3. Good health ,MESH: Cell Line ,MESH: DNA, Viral ,Biochemistry ,DNA, Viral ,biology.protein ,Origin recognition complex ,Primase ,Aptamers, Peptide ,Protein Binding - Abstract
International audience; 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.
- Published
- 2009