title: Sequence specificity for uridylylation of the viral peptide linked to the genome (VPg) of enteroviruses.
abstract: Enteroviruses (EV) uridylylate a peptide, VPg, as the first step in their replication. VPgpUpU, found free in infected cells, serves as the primer for RNA elongation. The abilities of four polymerases (3D(pol)), from EV-species A-C, to uridylylate VPgs that varied by up to 60% of their residues were compared. Each 3D(pol) was able to uridylylate all five VPgs using polyA RNA as template, while showing specificity for its own genome encoded peptide. All 3D(pol) uridylylated a consensus VPg representing the physical chemical properties of 31 different VPgs. Thus the residues required for uridylylation and the enzymatic mechanism must be similar in diverse EV. As VPg-binding sites differ in co-crystal structures, the reaction is probably done by a second 3D(pol) molecule. The conservation of polymerase residues whose mutation reduces uridylylation but not RNA elongation is compared.
authors.affiliation: Foundation for Applied Molecular Evolution (FfAME), 13709 Progress Blvd., Alachua, FL 32616, United States. Electronic address: chschein@ffame.org.
authors.forename: Catherine H
authors.initials: CH
authors.lastname: Schein
authors.affiliation: Dept. Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX 77555, United States.
authors.forename: Mengyi
authors.initials: M
authors.lastname: Ye
authors.affiliation: Dept. Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, NY 11790, United States.
authors.forename: Aniko V
authors.initials: AV
authors.lastname: Paul
authors.affiliation: Division of Viral Diseases, Centers for Disease Control and Prevention, 1600 Clifton Rd NE, MS G-17, Atlanta, GA 30333, United States.
authors.forename: M Steven
authors.initials: MS
authors.lastname: Oberste
authors.affiliation: Dept. Pathology and Microbiology, University of Nebraska Medical Center, NE 68198, United States.
authors.forename: Nora
authors.initials: N
authors.lastname: Chapman
authors.affiliation: Leiden Institute of Chemistry, Leiden University, P.O. Box 9502, 2300 RA, Leiden, The Netherlands.
authors.forename: Gerbrand J
authors.initials: GJ
authors.lastname: van der Heden van Noort
authors.affiliation: Leiden Institute of Chemistry, Leiden University, P.O. Box 9502, 2300 RA, Leiden, The Netherlands.
authors.forename: Dmitri V
authors.initials: DV
authors.lastname: Filippov
authors.affiliation: Dept. Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX 77555, United States.
authors.forename: Kyung H
authors.initials: KH
authors.lastname: Choi
mesh_terms: D004770:Enterovirus / Q000201:enzymology / Q000502:physiology*; D006801:Humans; D008958:Models, Molecular; D011487:Protein Conformation; D011499:Protein Processing, Post-Translational; D013379:Substrate Specificity; D014544:Uridine Triphosphate / Q000378:metabolism*; D017361:Viral Nonstructural Proteins / Q000737:chemistry / Q000378:metabolism*; D014779:Virus Replication
chemical_list: D017361:Viral Nonstructural Proteins; D014544:Uridine Triphosphate
keywords: Coxsackie virus; EV-71; Metal ion dependent phosphotransfer; Nucleotide transfer reaction; PCP-consensus sequence; Peptide priming; Poliovirus; RNA polymerase