title: Cotranslational coat protein-mediated inhibition of potyviral RNA translation.
abstract: Potato virus A (PVA) is a single-stranded positive-sense RNA virus and a member of the family Potyviridae. The PVA coat protein (CP) has an intrinsic capacity to self-assemble into filamentous virus-like particles, but the mechanism responsible for the initiation of viral RNA encapsidation in vivo remains unclear. Apart from virion assembly, PVA CP is also involved in the inhibition of viral RNA translation. In this study, we show that CP inhibits PVA RNA translation in a dose-dependent manner, through a mechanism involving the CP-encoding region. Analysis of this region, however, failed to identify any RNA secondary structure(s) preferentially recognized by CP, suggesting that the inhibition depends on CP-CP rather than CP-RNA interactions. In agreement with this possibility, insertion of an in-frame stop codon upstream of the CP sequence led to a marked decrease in the inhibition of viral RNA translation. Based on these results, we propose a model in which the cotranslational interactions between excess CP accumulating in trans and CP translated from viral RNA in cis are required to initiate the translational repression. This model suggests a mechanism for how viral RNA can be sequestered from translation and specifically selected for encapsidation at the late stages of viral infection.
  
  
  OBJECTIVE
  The main functions of the CP during potyvirus infection are to protect viral RNA from degradation and to transport it locally, systemically, and from host to host. Although virion assembly is a key step in the potyviral infectious cycle, little is known about how it is initiated and how viral RNA is selected for encapsidation. The results presented here suggest that CP-CP rather than CP-RNA interactions are predominantly involved in the sequestration of viral RNA away from translation. We propose that the cotranslational nature of these interactions may represent a mechanism for the selection of viral RNA for encapsidation. A better understanding of the mechanism of virion assembly may lead to development of crops resistant to potyviruses at the level of viral RNA encapsidation, thereby reducing the detrimental effects of potyvirus infections on food production.
authors: Department of Food and Environmental Sciences, University of Helsinki, Helsinki, Finland UMR 1332 Biologie du Fruit et Pathologie, INRA-Université Bordeaux 2, Villenave d'Ornon Cedex, France.
authors: Jane
authors: J
authors: Besong-Ndika
authors: Department of Food and Environmental Sciences, University of Helsinki, Helsinki, Finland.
authors: Konstantin I
authors: KI
authors: Ivanov
authors: Department of Food and Environmental Sciences, University of Helsinki, Helsinki, Finland.
authors: Anders
authors: A
authors: Hafrèn
authors: UMR 1332 Biologie du Fruit et Pathologie, INRA-Université Bordeaux 2, Villenave d'Ornon Cedex, France.
authors: Thierry
authors: T
authors: Michon
authors: Department of Food and Environmental Sciences, University of Helsinki, Helsinki, Finland kristiina.makinen@helsinki.fi.
authors: Kristiina
authors: http://orcid.org/0000-0001-8233-8942
authors: K
authors: Mäkinen
mesh_terms: D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism*; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D024202:Electrophoretic Mobility Shift Assay; D018274:Electroporation; D015967:Gene Expression Regulation, Viral / Q000235:genetics / Q000502:physiology*; D007163:Immunosorbent Techniques; D008854:Microscopy, Electron; D008957:Models, Genetic; D016296:Mutagenesis; D019176:Potyviridae / Q000235:genetics* / Q000378:metabolism; D014176:Protein Biosynthesis / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D014026:Nicotiana; D019065:Virus Assembly / Q000235:genetics / Q000502:physiology*
chemical_list: D036022:Capsid Proteins; D017931:DNA Primers; D018076:DNA, Complementary
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