Yeast two-hybrid study of tobacco rattle virus coat protein and 2b protein interactions
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The specificity of the interaction between the coat protein (CP) and 2b nematode-transmission helper protein of two isolates, PpK20 and PaY4, of Tobacco rattle virus (TRV) that differ in their transmission characteristics was investigated. Yeast two-hybrid experiments identified a central domain of the CP that promoted CP:CP interaction but inhibited CP:2b interaction. Deletion of a conserved coiled-coil domain from the 2b protein prevented its interaction with CP, whereas deletion of N- and C-terminal domains of the 2b protein greatly enhanced its interaction with CP. A C-terminal flexible domain of the CP was also shown to be important for interaction with the 2b protein. However, this domain was not sufficient to direct isolate-specific interaction of these proteins either in yeast or via a chimeric TRV in planta. Although these two TRV isolates are both transmitted by a shared vector trichodorid nematode, Paratrichodorus pachydermus, the CP from isolate PpK20 did not interact with the 2b protein from isolate PaY4, and vice versa, suggesting that in the field mixed infections of TRV are unlikely to cause promiscuous transmission by alternative trichodorid nematode species.
KeywordsTobacco Rattle Virus Scottish Crop Research Institute Produce Virus Particle Nematode Transmission Trichodorid Nematode
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- Karanastasi, I, Vassilakos, N, Roberts, IM, MacFarlane, SA, Brown, DJF 2000Immunogold localisation of tobacco rattle virus particles within the virus-vector nematode Paratrichodorus anemonesJ Nematol32512Google Scholar
- MacFarlane SA (1997) Natural recombination among plant virus genomes: evidence from tobraviruses. In: May MA (ed) Aspects of ssRNA virus genome organization. Sem Virol 8: 25–31, Academic Press, LondonGoogle Scholar
- Robinson, DJ 1994Sequences at the ends of RNA-2 of I6, a recombinant tobravirusArch Virol9245251Google Scholar
- Robinson, DJ, Harrison, BD 1985Unequal variation in the two genome parts of tobraviruses and evidence for the existence of three separate virusesJ Gen Virol66171176Google Scholar
- Vellios, EK, Brown, DJF, MacFarlane, SA 2002bSubstitution of a single amino acid in the PEBV 2b protein affects nematode transmissionJ Gen Virol8317711775Google Scholar
- Visser, PB, Bol, JF 1999Nonstructural proteins of Tobacco rattle virus which have a role in nematode-transmission: expression pattern and interaction with viral coat proteinJ Gen Virol8032723280Google Scholar