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Journal of Virology, June 2003, p. 7026-7033, Vol. 77, No. 12
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.12.7026-7033.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Functional Characterization of the Nuclear Localization Signal for a Suppressor of Posttranscriptional Gene Silencing

Xiangli Dong,1 Rene van Wezel,1 John Stanley,2 and Yiguo Hong1*

Horticulture Research International, East Malling, West Malling, Kent ME19 6BJ,1 Department of Disease and Stress Biology, John Innes Centre, Colney, Norwich NR4 7UH, United Kingdom2

Received 23 January 2003/ Accepted 28 March 2003

The nucleus-localized C2 protein of Tomato yellow leaf curl virus-China (TYLCV-C) is an active suppressor of posttranscriptional gene silencing (PTGS). Consistently, infection with TYLCV-C resulted in PTGS arrest in plants. The C2 protein possesses a functional, arginine-rich nuclear localization signal within the basic amino acid-rich region 17KVQHRIAKKTTRRRR31. When expressed from potato virus X, C2-RRRR31DVGG (in which the four consecutive arginine residues 28RRRR31 were replaced with DVGG) that had been tagged with a green fluorescent protein (GFP) failed to transport GFP into nuclei and was dysfunctional in inducing necrosis and suppressing PTGS in plants. Amino acid substitution mutants C2-K17D-GFP, C2-HR21DV-GFP, and C2-KK25DI-GFP localized to nuclei and produced necrosis, but only C2-K17D-GFP suppressed PTGS. The N-terminal portions C21-31 and C217-31 fused in frame to GFP were capable of targeting GFP to nuclei, but neither caused necrosis nor affected PTGS. Our data establish that nuclear localization is likely required for C2 protein to function in C2-mediated induction of necrosis and suppression of PTGS, which may follow diverse pathways in plants. Possible mechanisms of how the C2 protein involves these biological functions are discussed.

* Corresponding author. Mailing address: Horticulture Research International, East Malling, West Malling, Kent ME19 6BJ, United Kingdom. Phone: 44 173 284 3833. Fax: 44 173 284 9067. E-mail yiguo.hong{at}hri.ac.uk.

Journal of Virology, June 2003, p. 7026-7033, Vol. 77, No. 12
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.12.7026-7033.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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