HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Visser, P. B. Articles by Bol, J. F. Search for Related Content PubMed PubMed Citation Articles by Visser, P. B. Articles by Bol, J. F. Agricola Articles by Visser, P. B. Articles by Bol, J. F.

Nonstructural proteins of Tobacco rattle virus which have a role in nematode-transmission: expression pattern and interaction with viral coat protein

Institute of Molecular Plant Sciences, Gorlaeus Laboratories, PO Box 9502, 2300 RA Leiden, The Netherlands¹

Author for correspondence: John F. Bol. Fax +31 71 5274340. e-mail j.bol{at}chem.leidenuniv.nl

RNA 2 of Tobacco rattle virus isolate PpK20 encodes the viral coat protein (CP) and two nonstructural proteins of 40 kDa (‘40K protein’) and 32·8 kDa (‘32·8K’). The 40K protein is required for transmission of the virus by the vector nematode Paratrichodorus pachydermus whereas the 32·8K protein may be involved in transmission by other vector nematode species. An antiserum was raised against the 40K protein expressed in E. coli and used to study the expression and subcellular localization of this protein in infected Nicotiana benthamiana plants. The time-course of the expression of the 40K protein in leaves and roots was similar to that of CP and both proteins were similarly distributed over the 1000 g pellet, 30000 g pellet and 30000 g supernatant fractions of leaf and root homogenates. Using the yeast two-hybrid system, a strong interaction between CP subunits was observed and weaker interactions between CP and the 32·8K protein and between CP and the 40K protein were detected. A deletion of the C-terminal 19 amino acids of CP interfered with the CP–40K interaction but not with CP–32·8K or CP–CP interactions, whereas a C-terminal deletion of 79 amino acids interfered with CP–40K and CP–32·8K interactions but not with the CP–CP interaction. As the C terminus of CP is known to be involved in nematode-transmission of tobraviruses, the data support the hypothesis that interactions between CP and RNA 2-encoded nonstructural proteins play a role in the transmission process.