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Journal of Virology, June 2005, p. 7217-7226, Vol. 79, No. 11
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.11.7217-7226.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Aureusvirus P14 Is an Efficient RNA Silencing Suppressor That Binds Double-Stranded RNAs without Size Specificity{ddagger}

Zsuzsanna Mérai,1 Zoltán Kerényi,1 Attila Molnár,1,{dagger} Endre Barta,1 Anna Válóczi,1 György Bisztray,2 Zoltán Havelda,1 József Burgyán,1 and Dániel Silhavy1*

Agricultural Biotechnology Center, Gödöllö, Hungary,1 Department of Genetics and Horticultural Plant Breeding, Budapest University of Economic Sciences and Public Administration, Budapest, Hungary2

Received 1 September 2004/ Accepted 17 January 2005

RNA silencing is a conserved eukaryotic gene regulatory system in which sequence specificity is determined by small RNAs. Plant RNA silencing also acts as an antiviral mechanism; therefore, viral infection requires expression of a silencing suppressor. The mechanism and the evolution of silencing suppression are still poorly understood. Tombusvirus open reading frame (ORF) 5-encoded P19 is a size-selective double-stranded RNA (dsRNA) binding protein that suppresses silencing by sequestering double-stranded small interfering RNAs (siRNAs), the specificity determinant of the antiviral silencing system. To better understand the evolution of silencing suppression, we characterized the suppressor of the type member of Aureusviruses, the closest relatives of the genus Tombusvirus. We show that the Pothos latent virus (PoLV) ORF 5-encoded P14 is an efficient suppressor of both virus- and transgene-induced silencing. Findings that in vitro P14 binds dsRNAs and double-stranded siRNAs without obvious size selection suggest that P14, unlike P19, can suppress silencing by sequestering both long dsRNA and double-stranded siRNA components of the silencing machinery. Indeed, P14 prevents the accumulation of hairpin transcript-derived siRNAs, indicating that P14 inhibits inverted repeat-induced silencing by binding the long dsRNA precursors of siRNAs. However, viral siRNAs accumulate to high levels in PoLV-infected plants; therefore, P14 might inhibit virus-induced silencing by sequestering double-stranded siRNAs. Finally, sequence analyses suggest that P14 and P19 suppressors diverged from an ancient dsRNA binding suppressor that evolved as a nested protein within the common ancestor of aureusvirus-tombusvirus movement proteins.

* Corresponding author. Mailing address: Agricultural Biotechnology Center, Plant Science Institute, P.O. Box 411, H-2101 Gödöllö, Hungary. Phone: 36 28 526 194. Fax: 36 28 526 145. E-mail: silhavy{at}abc.hu.

{ddagger} Supplemental material for this article may be found at http://jvi.asm.org/.

{dagger} Present address: The Sainsbury Laboratory, John Innes Centre, Colney Lane, Norwich NR4 7UH, United Kingdom.

Journal of Virology, June 2005, p. 7217-7226, Vol. 79, No. 11
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.11.7217-7226.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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