title: Involvement of a joker mutation in a polymerase-independent lethal mutagenesis escape mechanism.
abstract: We previously characterized a foot-and-mouth disease virus (FMDV) with three amino acid replacements in its polymerase (3D) that conferred resistance to the mutagenic nucleoside analogue ribavirin. Here we show that passage of this mutant in the presence of high ribavirin concentrations resulted in selection of viruses with the additional replacement I248T in 2C. This 2C substitution alone (even in the absence of replacements in 3D) increased FMDV fitness mainly in the presence of ribavirin, prevented an incorporation bias in favor of A and U associated with ribavirin mutagenesis, and conferred the ATPase activity of 2C decreased sensitivity to ribavirin-triphosphate. Since in previous studies we described that 2C with I248T was selected under different selective pressures, this replacement qualifies as a joker substitution in FMDV evolution. The results have identified a role of 2C in nucleotide incorporation, and have unveiled a new polymerase-independent mechanism of virus escape to lethal mutagenesis.
authors.affiliation: Centro de Biologia Molecular "Severo Ochoa" (CSIC-UAM), Cantoblanco, E-28049 Madrid, Spain.
authors.forename: Rubén
authors.initials: R
authors.lastname: Agudo
authors.affiliation: Centro de Biologia Molecular "Severo Ochoa" (CSIC-UAM), Cantoblanco, E-28049 Madrid, Spain.
authors.forename: Ignacio
authors.initials: I
authors.lastname: de la Higuera
authors.affiliation: Centro de Biologia Molecular "Severo Ochoa" (CSIC-UAM), Cantoblanco, E-28049 Madrid, Spain.
authors.forename: Armando
authors.initials: A
authors.lastname: Arias
authors.affiliation: Instituto de Hortofruticultura Subtropical y Mediterránea "La Mayora", Universidad de Málaga - Consejo Superior de Investigaciones Científicas, (IHSM-UMA-CSIC) Área de Genética, Campus de Teatinos, 29071 Málaga, Spain.
authors.forename: Ana
authors.initials: A
authors.lastname: Grande-Pérez
authors.affiliation: Centro de Biologia Molecular "Severo Ochoa" (CSIC-UAM), Cantoblanco, E-28049 Madrid, Spain; Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Barcelona, Spain. Electronic address: edomingo@cbm.csic.es.
authors.forename: Esteban
authors.initials: E
authors.lastname: Domingo
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism; D000956:Antigens, Viral / Q000235:genetics / Q000378:metabolism; D000998:Antiviral Agents / Q000494:pharmacology; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D024882:Drug Resistance, Viral; D004789:Enzyme Activation; D029544:Foot-and-Mouth Disease Virus / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D007700:Kinetics; D050296:Microbial Viability / Q000187:drug effects / Q000235:genetics*; D009154:Mutation; D012254:Ribavirin / Q000494:pharmacology; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D000956:Antigens, Viral; D000998:Antiviral Agents; D002352:Carrier Proteins; D017361:Viral Nonstructural Proteins; C103557:virus-infection-associated antigen, Foot-and-mouth disease virus; D012254:Ribavirin; D012321:DNA-Directed RNA Polymerases; C076023:2C protein, viral; D000251:Adenosine Triphosphatases
keywords: Error catastrophe; Foot-and-mouth disease virus; Quasispecies; Ribavirin; Virus extinction