title: Low-Fidelity Polymerases of Alphaviruses Recombine at Higher Rates To Overproduce Defective Interfering Particles.
abstract: Low-fidelity RNA-dependent RNA polymerases for many RNA virus mutators have been shown to confer attenuated phenotypes, presumably due to increased mutation rates. Additionally, for many RNA viruses, replication to high titers results in the production of defective interfering particles (DIs) that also attenuate infection. We hypothesized that fidelity, recombination, and DI production are tightly linked. We show that a Sindbis virus mutator replicating at a high multiplicity of infection manifests an earlier and greater accumulation of DIs than its wild-type counterpart. The isolated DIs interfere with the replication of full-length virus in a dose-dependent manner. Importantly, the ability of the mutator virus to overproduce DIs could be linked to an increased recombination frequency. These data confirm that RNA-dependent RNA polymerase fidelity and recombination are inversely correlated for this mutator. Our findings suggest that defective interference resulting from higher recombination rates may be more detrimental to RNA virus mutators than the increase in mutational burden.
  
  
  OBJECTIVE
  Replication, adaptation, and evolution of RNA viruses rely in large part on their low-fidelity RNA-dependent RNA polymerase. Viruses artificially modified in their polymerases to decrease fidelity (mutator viruses) are attenuated in vivo, demonstrating the important role of fidelity in viral fitness. However, attenuation was attributed solely to the modification of the viral mutation rate and the accumulation of detrimental point mutations. In this work, we described an additional phenotype of mutator viruses: an increased recombination rate leading to defective interfering particle (DI) overproduction. Because DIs are known for their inhibitory effect on viral replication, our work suggests that fidelity variants may be attenuated in vivo via several mechanisms. This has important implications in the development of fidelity variants as live attenuated vaccine strains.
authors.affiliation: Institut Pasteur, Centre National de la Recherche Scientifique, UMR 3569, Paris, France University of Paris Diderot, Sorbonne Paris Cité, Cellule Pasteur, Paris, France.
authors.forename: Enzo Z
authors.initials: EZ
authors.lastname: Poirier
authors.affiliation: Institut Pasteur, Centre National de la Recherche Scientifique, UMR 3569, Paris, France.
authors.forename: Bryan C
authors.initials: BC
authors.lastname: Mounce
authors.affiliation: Institut Pasteur, Centre National de la Recherche Scientifique, UMR 3569, Paris, France University of Paris Diderot, Sorbonne Paris Cité, Cellule Pasteur, Paris, France.
authors.forename: Kathryn
authors.initials: K
authors.lastname: Rozen-Gagnon
authors.affiliation: Institut Pasteur, Centre National de la Recherche Scientifique, UMR 3569, Paris, France.
authors.forename: Peter Jan
authors.initials: PJ
authors.lastname: Hooikaas
authors.affiliation: Institut Pasteur, Centre National de la Recherche Scientifique, UMR 3569, Paris, France.
authors.forename: Kenneth A
authors.initials: KA
authors.lastname: Stapleford
authors.affiliation: Institut Pasteur, Centre National de la Recherche Scientifique, UMR 3569, Paris, France.
authors.forename: Gonzalo
authors.initials: G
authors.lastname: Moratorio
authors.affiliation: Institut Pasteur, Centre National de la Recherche Scientifique, UMR 3569, Paris, France marco.vignuzzi@pasteur.fr.
authors.forename: Marco
authors.initials: M
authors.lastname: Vignuzzi
mesh_terms: D000818:Animals; D002460:Cell Line; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D003673:Defective Viruses / Q000235:genetics / Q000254:growth & development*; D012367:RNA, Viral / Q000096:biosynthesis*; D012845:Sindbis Virus / Q000201:enzymology* / Q000235:genetics; D014762:Viral Interference; D014779:Virus Replication
chemical_list: D012367:RNA, Viral; D012321:DNA-Directed RNA Polymerases