A strategy for screening an inhibitor of viral silencing suppressors, which attenuates symptom development of plant viruses

Shimura, Hanako; Fukagawa, Takako; Meguro, Ayano; Yamada, Hirokazu; Oh-hira, Mahito; Sano, Shinsuke; Masuta, Chikara

doi:10.1016/j.febslet.2008.10.046

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Volume 582, Issue 29 , Pages 4047-4052, 10 December 2008

A strategy for screening an inhibitor of viral silencing suppressors, which attenuates symptom development of plant viruses

Edited by Shou-Wei Ding

Hanako Shimura
- Laboratory of Cell Biology and Manipulation, Graduate School of Agriculture, Hokkaido University, Sapporo 060-8589, Japan
Takako Fukagawa
- Odawara Research Center, Nippon Soda Co., Ltd., 345, Takada Odawara, Kanagawa 250-0280, Japan
Ayano Meguro
- Laboratory of Cell Biology and Manipulation, Graduate School of Agriculture, Hokkaido University, Sapporo 060-8589, Japan
Hirokazu Yamada
- Odawara Research Center, Nippon Soda Co., Ltd., 345, Takada Odawara, Kanagawa 250-0280, Japan
Mahito Oh-hira
- Odawara Research Center, Nippon Soda Co., Ltd., 345, Takada Odawara, Kanagawa 250-0280, Japan
Shinsuke Sano
- Odawara Research Center, Nippon Soda Co., Ltd., 345, Takada Odawara, Kanagawa 250-0280, Japan
Chikara Masuta
- Laboratory of Cell Biology and Manipulation, Graduate School of Agriculture, Hokkaido University, Sapporo 060-8589, Japan
- Corresponding author. Fax: +81 11 7062483.

Received 4 September 2008; received in revised form 16 October 2008; accepted 31 October 2008. published online 07 November 2008.

Abstract

To find out whether we can control plant virus diseases by blocking viral RNA silencing suppressors (RSSs), we developed a strategy to screen inhibitors that block the association of RSSs with siRNAs using a surface plasmon resonance assay. The screened chemicals were tested in competition with RSSs for binding to siRNAs using a mobility shift assay. We then confirmed that tested chemicals actually inhibited the RSS activity in vivo using a protoplast assay which was developed for this purpose. This entire system can be adapted to screening inhibitors of not only plant viruses but also some animal viruses possessing RSSs.

Keywords: Double-stranded RNA, RNA silencing suppressor, Virus, Inhibitor