title: Amiloride inhibits the initiation of Coxsackievirus and poliovirus RNA replication by inhibiting VPg uridylylation.
abstract: The mechanism of amiloride inhibition of Coxsackievirus B3 (CVB3) and poliovirus type 1 (PV1) RNA replication was investigated using membrane-associated RNA replication complexes. Amiloride was shown to inhibit viral RNA replication and VPgpUpU synthesis. However, the drug had no effect on polymerase elongation activity during either (-) strand or (+) strand synthesis. These findings indicated that amiloride inhibited the initiation of RNA synthesis by inhibiting VPg uridylylation. In addition, in silico binding studies showed that amiloride docks in the VPg binding site on the back of the viral RNA polymerase, 3D(pol). Since VPg binding at this site on PV1 3D(pol) was previously shown to be required for VPg uridylylation, our results suggest that amiloride inhibits VPg binding to 3D(pol). In summary, our findings are consistent with a model in which amiloride inhibits VPgpUpU synthesis and viral RNA replication by competing with VPg for binding to 3D(pol).
authors: Department of Biochemistry and Molecular Biology, University of Florida, College of Medicine, Gainesville, FL 32610-0245, USA.
authors: Sushma A
authors: SA
authors: Ogram
authors: Department of Biochemistry and Molecular Biology, University of Florida, College of Medicine, Gainesville, FL 32610-0245, USA.
authors: Christopher D
authors: CD
authors: Boone
authors: Department of Biochemistry and Molecular Biology, University of Florida, College of Medicine, Gainesville, FL 32610-0245, USA.
authors: Robert
authors: R
authors: McKenna
authors: Department of Biochemistry and Molecular Biology, University of Florida, College of Medicine, Gainesville, FL 32610-0245, USA. Electronic address: Flanegan@ufl.edu.
authors: James B
authors: JB
authors: Flanegan
mesh_terms: D000584:Amiloride / Q000737:chemistry / Q000494:pharmacology*; D000998:Antiviral Agents / Q000737:chemistry / Q000494:pharmacology*; D003384:Coxsackievirus Infections / Q000821:virology; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015536:Down-Regulation / Q000187:drug effects; D004770:Enterovirus / Q000737:chemistry / Q000187:drug effects* / Q000235:genetics* / Q000502:physiology; D006801:Humans; D008958:Models, Molecular; D011051:Poliomyelitis / Q000821:virology; D017955:Poliovirus / Q000737:chemistry / Q000187:drug effects* / Q000235:genetics* / Q000502:physiology; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D012367:RNA, Viral / Q000235:genetics* / Q000378:metabolism; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014779:Virus Replication / Q000187:drug effects*
chemical_list: D000998:Antiviral Agents; D012367:RNA, Viral; D014764:Viral Proteins; D000584:Amiloride; D012321:DNA-Directed RNA Polymerases
keywords: (+) strand RNA; (−) strand RNA; Amiloride; Coxsackievirus B3 (CVB3); Poliovirus (PV); RNA replication complex; VPg; VPg uridylylation