title: An interfering activity against lymphocytic choriomeningitis virus replication associated with enhanced mutagenesis.
abstract: Previous studies have documented that, in the presence of the mutagenic base analogue 5-fluorouracil (FU), lymphocytic choriomeningitis virus (LCMV) that persisted in BHK-21 cells decreased its infectivity to a larger extent than intracellular viral RNA levels, prior to virus extinction. This observation, together with in silico simulations, led to the proposal of the lethal defection model of virus extinction. This model suggests the participation of defective-interfering genomes in the loss of infectivity by increased mutagenesis. Since LCMV naturally produces defective-interfering particles, it was important to show that a capacity to interfere is produced in association with FU treatment. Here, we document that BHK-21 cells persistently infected with LCMV grown in the presence of FU, but not in its absence, generated an interfering activity that suppressed LCMV infectivity. Interference was specific for LCMV and was sensitive to UV irradiation and its activity was dose- and time-dependent. The interfering preparations produced positive LCMV immunofluorescence and viral particles seen by electron microscopy when used to infect cells, despite some preparations being devoid of detectable infectivity. Interference did not involve significant increases of mutant spectrum complexity, as predicted by the lethal defection model. The results provide support for a specific interference associated with LCMV when the virus replicates in the presence of FU. The excess of interference relative to that observed in the absence of FU is necessary for LCMV extinction.
authors.affiliation: Centro de Biología Molecular 'Severo Ochoa' (CSIC-UAM), C/ Nicolás Cabrera, 1, Universidad Autónoma de Madrid, Cantoblanco, 28049 Madrid, Spain.
authors.forename: Verónica
authors.initials: V
authors.lastname: Martín
authors.forename: David
authors.initials: D
authors.lastname: Abia
authors.forename: Esteban
authors.initials: E
authors.lastname: Domingo
authors.forename: Ana
authors.initials: A
authors.lastname: Grande-Pérez
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000032:analysis; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D005472:Fluorouracil / Q000494:pharmacology; D008217:Lymphocytic choriomeningitis virus / Q000235:genetics / Q000502:physiology*; D016296:Mutagenesis; D014466:Ultraviolet Rays; D014709:Vero Cells; D014762:Viral Interference; D014771:Virion / Q000302:isolation & purification; D014779:Virus Replication
chemical_list: D000956:Antigens, Viral; D005472:Fluorouracil