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Originally published as JGV in Press, 10.1099/vir.0.010488-0 on March 12, 2009 J Gen Virol 90 (2009), 1762-1766; DOI 10.1099/vir.0.010488-0

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Short Communication

Binding of small interfering RNA molecules is crucial for RNA interference suppressor activity of rice hoja blanca virus NS3 in plants

Hans Hemmes{ddagger}, Lucas Kaaij§, Dick Lohuis, Marcel Prins||, Rob Goldbach{dagger} and Esther Schnettler

Laboratory of Virology, Wageningen University, Binnenhaven 11, 6709 PD Wageningen, The Netherlands

Correspondence
Esther Schnettler
Esther.Schnettler{at}wur.nl
Hans Hemmes
jhemmes{at}mail.rockefeller.edu

The NS3 protein of rice hoja blanca virus represents a viral suppressor of RNA interference (RNAi) that sequesters small interfering (si)RNAs in vitro. To determine whether this siRNA binding property is the critical determinant for the suppressor activity of NS3, NS3 was altered by alanine point mutations and the resulting mutant proteins were tested for both siRNA binding ability and RNAi suppressor activity in plants. Alanine substitutions of lysine residues at positions 173–175 resulted in mutant proteins that lost both their affinity for siRNAs and their RNAi suppressor activity in planta. This indicates that siRNA binding of NS3 is indeed essential for the suppressor function of NS3 and that residues at positions 173–175 are involved in the siRNA binding and suppressor activities.

{dagger}Dedicated to Rob Goldbach who sadly passed away on 7 April 2009.

{ddagger}Present address: Rockefeller University, Laboratory of Plant Molecular Biology, 1230 York Avenue, New York, NY 10065, USA.

§Present address: Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences & University Medical Centre Utrecht, Uppsalalaan 8, 3584 CT Utrecht, The Netherlands.

||Present address: Keygene N.V., Agro Business Park 90, 6708 PW Wageningen, The Netherlands.






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