Articles
THRIPS MIDGUT PROTEINS THAT BIND TO TOMATO SPOTTED WILT VIRUS GLYCOPROTEINS
Article number
431_29
Pages
341 – 349
Language
Abstract
Electron microscopy studies indicate that tomato spotted wilt virus (TSWV) uptake in western flower thrips (WFT) (Frankliniella occidentalis) is receptor-mediated.
The location of the viral glycoproteins (GP) in the virion envelope and the presence of a cell-attachment motif in the G2 glycoprotein, supports the involvement of the viral glycoproteins in the process of virus uptake.
Whole insects and dissected midguts of WFT were subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), the proteins transferred to nitrocellulose, and then the blot probed with isolated TSWV, macerated TSWV infected tissue, or a GP fraction from isolated virus.
This was followed by antibodies to selected TSWV proteins.
A protein of about 50 kDa was detected using isolated virus to first probe the WFT blots followed by a murine antibody to the GP fraction of TSWV, but not with antibody to the nonstructural protein encoded by the small RNA (NSs) or nucleocapsid (N) proteins of TSWV. Preparations from either Circulifer tenellus or Schizaphis graminum were not labeled in this manner.
When a biotin-avidin system was used to enhance detection of GP binding to thrips proteins, proteins of 50 kDa (a doublet) and 38 kDa were detected.
The relevance of these findings to thrips acquisition of TSWV is discussed.
The location of the viral glycoproteins (GP) in the virion envelope and the presence of a cell-attachment motif in the G2 glycoprotein, supports the involvement of the viral glycoproteins in the process of virus uptake.
Whole insects and dissected midguts of WFT were subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), the proteins transferred to nitrocellulose, and then the blot probed with isolated TSWV, macerated TSWV infected tissue, or a GP fraction from isolated virus.
This was followed by antibodies to selected TSWV proteins.
A protein of about 50 kDa was detected using isolated virus to first probe the WFT blots followed by a murine antibody to the GP fraction of TSWV, but not with antibody to the nonstructural protein encoded by the small RNA (NSs) or nucleocapsid (N) proteins of TSWV. Preparations from either Circulifer tenellus or Schizaphis graminum were not labeled in this manner.
When a biotin-avidin system was used to enhance detection of GP binding to thrips proteins, proteins of 50 kDa (a doublet) and 38 kDa were detected.
The relevance of these findings to thrips acquisition of TSWV is discussed.
Authors
M.D. Bandla, D.M. Westcot, D.E. Ullman, J.L. Sherwood
Keywords
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