Structure of Foot-and-Mouth Disease Virus Mutant Polymerases with Reduced Sensitivity to Ribavirin

• DOI: 10.1128/jvi.02420-09
• Publication Date: 2010-04-15T01:34:58Z
• Authors: Cristina Ferrer-Orta; Macarena Sierra; Rubén Agudo; Ignacio de la Hig...; Armando Arias; Rosa Pérez-Luque; Cristina Escarmís; Esteban Domingo; Nuria Verdaguer
• Keywords: Picornavirus

ABSTRACT Passage of poliovirus (PV) or foot-and-mouth disease virus (FMDV) in the presence ribavirin (R) selected for viruses with decreased sensitivity to R, which included different mutations their polymerase (3D): G64S located finger subdomain case PV and M296I within loop β9-α11 at active site FMDV. To investigate why disparate substitutions were two closely related 3Ds, we constructed FMDVs a 3D that either G62S (the equivalent replacement FMDV G64S), M296I, both substitutions. G62S, but not inflicts upon strong selective disadvantage is partially compensated by substitution M296I. The corresponding mutant polymerases, 3D(G62S), 3D(M296I), 3D(G62S-M296I), analyzed functionally structurally. impairs RNA-binding, polymerization, R monophosphate incorporation activities. X-ray structures 3D(G62S)-RNA, 3D(M296I)-RNA, 3D(G62S-M296I)-RNA complexes show although positions are separated 13.1 Å, loops where replacements reside tightly connected through an extensive network interactions reach site. In particular, seems restrict flexibility and, as consequence, its ability bind RNA template. Thus, localized change may affect catalytic domain. results provide structural interpretation amino acid confer resistance reveal complex intra-3D can recognition template incoming nucleotide.