Functional characterization of the Beet necrotic yellow vein virus RNA-5-encoded p26 protein: evidence for structural pathogenicity determinants
| Autor: | Didier Link, Angèle Geldreich, Elodie Klein, David Gilmer, Olivier Lemaire, Laure Schmidlin, Audrey Schirmer, Mathieu Erhardt |
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| Rok vydání: | 2005 |
| Předmět: |
Gene Expression Regulation
Viral Recombinant Fusion Proteins animal diseases Green Fluorescent Proteins Molecular Sequence Data Biology Benyvirus Virus Plant Viruses Viral Proteins Transcription (biology) Virology RNA Viruses Beet necrotic yellow vein virus Amino Acid Sequence Replicon Chenopodium quinoa Nuclear export signal Plant Diseases Reporter gene Base Sequence RNA biology.organism_classification Molecular biology Plant Leaves RNA Viral Beta vulgaris |
| Zdroj: | Journal of General Virology. 86:2115-2125 |
| ISSN: | 1465-2099 0022-1317 |
| DOI: | 10.1099/vir.0.80937-0 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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