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J Gen Virol 86 (2005), 2115-2125; DOI 10.1099/vir.0.80937-0

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© 2005 Society for General Microbiology

Functional characterization of the Beet necrotic yellow vein virus RNA-5-encoded p26 protein: evidence for structural pathogenicity determinants

Didier Link1, Laure Schmidlin1, Audrey Schirmer2, Elodie Klein1, Mathieu Erhardt1, Angèle Geldreich1, Olivier Lemaire2 and David Gilmer1

1 Institut de Biologie Moléculaire des Plantes, 12 rue du Général Zimmer, 67084 Strasbourg cedex, France
2 INRA, UR-BIVV, 28 rue de Herrlisheim, 68021 Colmar, France

Correspondence
David Gilmer
david.gilmer{at}ibmp-ulp.u-strasbg.fr

A Beet necrotic yellow vein virus isolate containing a fifth RNA is present in the Pithiviers area of France. A full-length cDNA clone of RNA-5 was obtained and placed under the control of a T7-RNA-pol promoter that allowed the production of infectious transcripts. ‘Pithiviers' isolate-specific necrotic symptoms were obtained on Chenopodium quinoa when RNA-5-encoded p26 was expressed either from RNA-5 or from an RNA-3-derived replicon. By using haemagglutinin- and green fluorescent protein-tagged constructs, virally expressed p26-fusion proteins induced the same necrotic local lesions on host plants and were localized mainly in the nucleus of infected cells. Deletion mutagenesis permitted identification of two domains, responsible respectively for nuclear export and cytoplasmic retention of the p26 mutated proteins. By using a yeast two-hybrid system, Gal4DB–p26 protein self-activated transcription of the His3 reporter gene. The p26 transcription-activation domain was located within its first 55 aa and has been studied by alanine scanning. Resulting p26 mutants were tested for their capability to induce necrotic symptoms and to localize in the nuclear compartment.

The GenBank/EMBL/DDBJ accession number for the full-length infectious transcript of RNA-5 described in this work is AY823407.




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A. Schirmer, D. Link, V. Cognat, B. Moury, M. Beuve, A. Meunier, C. Bragard, D. Gilmer, and O. Lemaire
Phylogenetic analysis of isolates of Beet necrotic yellow vein virus collected worldwide
J. Gen. Virol., October 1, 2005; 86(10): 2897 - 2911.
[Abstract] [Full Text] [PDF]


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