title: Allosteric inhibitors of Coxsackie virus A24 RNA polymerase.
abstract: Coxsackie virus A24 (CVA24), a causative agent of acute hemorrhagic conjunctivitis, is a prototype of enterovirus (EV) species C. The RNA polymerase (3D(pol)) of CVA24 can uridylylate the viral peptide linked to the genome (VPg) from distantly related EV and is thus, a good model for studying this reaction. Once UMP is bound, VPgpU primes RNA elongation. Structural and mutation data have identified a conserved binding surface for VPg on the RNA polymerase (3D(pol)), located about 20Å from the active site. Here, computational docking of over 60,000 small compounds was used to select those with the lowest (best) specific binding energies (BE) for this allosteric site. Compounds with varying structures and low BE were assayed for their effect on formation of VPgU by CVA24-3D(pol). Two compounds with the lowest specific BE for the site inhibited both uridylylation and formation of VPgpolyU at 10-20μM. These small molecules can be used to probe the role of this allosteric site in polymerase function, and may be the basis for novel antiviral compounds.
authors.affiliation: Foundation for Applied Molecular Evolution, 13709 Progress Blvd, Box 7, Alachua, FL 32616, United States; Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX 77555, United States. Electronic address: chschein@ffame.org.
authors.forename: Catherine H
authors.initials: CH
authors.lastname: Schein
authors.affiliation: Foundation for Applied Molecular Evolution, 13709 Progress Blvd, Box 7, Alachua, FL 32616, United States.
authors.forename: Diane
authors.initials: D
authors.lastname: Rowold
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX 77555, United States; Sealy Center for Structural Biology and Molecular Biophysics, UTMB, United States.
authors.forename: Kyung H
authors.initials: KH
authors.lastname: Choi
mesh_terms: D000494:Allosteric Regulation / Q000187:drug effects; D000998:Antiviral Agents / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D012321:DNA-Directed RNA Polymerases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D030015:Enterovirus C, Human / Q000187:drug effects* / Q000201:enzymology; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D008826:Microbial Sensitivity Tests; D062105:Molecular Docking Simulation; D015394:Molecular Structure; D054852:Small Molecule Libraries / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D014779:Virus Replication / Q000187:drug effects
chemical_list: D000998:Antiviral Agents; D004791:Enzyme Inhibitors; D054852:Small Molecule Libraries; D012321:DNA-Directed RNA Polymerases
keywords: +-Strand RNA virus; Antiviral compounds; Coxsackie virus; Docking compound libraries; Enterovirus RNA polymerase; Poliovirus; Protein-primed RNA synthesis; Uridylylation mechanism