title: Rapid screening for dominant negative mutations in the beet necrotic yellow vein virus triple gene block proteins P13 and P15 using a viral replicon.
abstract: Point mutations were introduced into the genes encoding the triple gene bock movement proteins P13 and P15 of beet necrotic yellow vein virus (BNYVV). Mutations which disabled viral cell-to-cell movement in Chenopodium quinoa were then tested for their ability to act as dominant negative inhibiters of movement of wild-type BNYVV when expressed from a co-inoculated BNYVV RNA 3-based replicon. For P13, three types of mutation inhibited the movement function: non-synomynous mutations in the N- and C-terminal hydrophobic domains, a mutation at the boundary between the N-terminal hydrophobic domain and the central hydrophilic domain (mutant P13-A12), and mutations in the conserved sequence motif in the central hydrophilic domain. However, only the 'boundary' mutant P13-A12 strongly inhibited movement of wild-type virus when expressed from the co-inoculated replicon. Similar experiments with P15 detected four movement-defective mutants which strongly inhibited cell-to-cell movement of wild-type BNYVV when the mutants were expressed from a co-inoculated replicon. Beta vulgaris transformed with two of these P15 mutants were highly resistant to fungus-mediated infection with BNYVV.
authors.affiliation: Institut de Biologie Moléculaire des Plantes du CNRS et de l'Université Louis Pasteur, Strasbourg, France.
authors.forename: E
authors.initials: E
authors.lastname: Lauber
authors.forename: L
authors.initials: L
authors.lastname: Janssens
authors.forename: G
authors.initials: G
authors.lastname: Weyens
authors.forename: G
authors.initials: G
authors.lastname: Jonard
authors.forename: K E
authors.initials: KE
authors.lastname: Richards
authors.forename: M
authors.initials: M
authors.lastname: Lefèbvre
authors.forename: H
authors.initials: H
authors.lastname: Guilley
mesh_terms: D005658:Fungi / Q000502:physiology; D005799:Genes, Dominant; D005814:Genes, Viral / Q000235:genetics*; D009068:Movement; D018515:Plant Leaves / Q000235:genetics*; D010942:Plant Viruses / Q000235:genetics*; D030821:Plants, Genetically Modified; D017354:Point Mutation; D012328:RNA Viruses / Q000502:physiology*; D012367:RNA, Viral / Q000096:biosynthesis* / Q000235:genetics; D012093:Replicon / Q000235:genetics; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000235:genetics*
chemical_list: D012367:RNA, Viral; D014764:Viral Proteins