Approaches for the Generation of New Anti-cytomegalovirus Agents: Identification of Protein–Protein Interaction Inhibitors and Compounds Against the HCMV IE2 Protein
- 1.9k Downloads
Human cytomegalovirus (HCMV) infection is responsible for severe, often even fatal, diseases in immunocompromised subjects and also represents the major cause of viral-associated congenital malformations in newborn children. The few drugs licensed for anti-HCMV therapy suffer from many drawbacks and none of them have been approved for the treatment of congenital infections. Furthermore, the emergence of drug-resistant viral strains represents a major concern for disease management. Thus, there is a strong need for new anti-HCMV drugs. Here we describe three different assays for the discovery of novel anti-HCMV compounds: two are in vitro assays, i.e., a fluorescence polarization (FP)-based assay and an enzyme-linked immunosorbent assay (ELISA), which are designed to search for compounds that act by disrupting the interactions between the HCMV DNA polymerase subunits, but in general can be employed to find inhibitors of any protein–protein interaction of interest; the third is a cell-based assay designed to identify inhibitors of the viral immediate-early 2 (IE2) protein activities.
Key wordsAntivirals Cell-based assay DNA polymerase inhibitors Enzyme-linked immunosorbent assay Fluorescence polarization Human cytomegalovirus IE2 protein Protein–protein interaction
The research in our laboratories was supported by MURST EX60%, Progetto di Ricerca di Ateneo 2007 (grant no. CPDA074945), and PRIN 2008 (grant no. 20085FF4J4) to A.L., by PRIN and the Piedmont Region (Ricerca Sanitaria Finalizzata) to G.G., and by Regione Veneto and Progetto Strategico di Ateneo 2008 to G.P.
- 7.Asmar J, Wiebusch L, Truss M, Hagemeier C (2004) The putative zinc finger of the human cytomegalovirus IE2 86-kilodalton protein is dispensable for DNA binding and autorepression, thereby demarcating a concise core domain in the C-terminus of the protein. J Virol 78:11853–11864PubMedCentralPubMedCrossRefGoogle Scholar
- 10.Loregian A, Mercorelli B, Muratore G, Sinigalia E, Massari S, Gribaudo G, Gatto B, Tabarrini O, Palumbo M, Cecchetti V, Palù G (2010) The 6-aminoquinolone WC5 inhibits human cytomegalovirus replication at an early stage by interfering with the transactivating activity of viral immediate-early 2 protein. Antimicrob Agents Chemother 54:1930–1940PubMedCentralPubMedCrossRefGoogle Scholar