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Mutational analysis of two highly conserved motifs in the silencing suppressor encoded by tomato spotted wilt virus (genus Tospovirus, family Bunyaviridae)

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Abstract

Tospoviruses cause serious economic losses to a wide range of field and horticultural crops on a global scale. The NSs gene encoded by tospoviruses acts as a suppressor of host plant defense. We identified amino acid motifs that are conserved in all of the NSs proteins of tospoviruses for which the sequence is known. Using tomato spotted wilt virus (TSWV) as a model, the role of these motifs in suppressor activity of NSs was investigated. Using site-directed point mutations in two conserved motifs, glycine, lysine and valine/threonine (GKV/T) at positions 181-183 and tyrosine and leucine (YL) at positions 412-413, and an assay to measure the reversal of gene silencing in Nicotiana benthamiana line 16c, we show that substitutions (K182 to A, and L413 to A) in these motifs abolished suppressor activity of the NSs protein, indicating that these two motifs are essential for the RNAi suppressor function of tospoviruses.

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Acknowledgements

This work was supported by the WSU Agricultural Research Center, PPNS No. 0627, Department of Plant Pathology, College of Agricultural, Human and Natural Resource Sciences, Agricultural Research Center, Project # WNPO 0545, Washington State University, Pullman, WA, 99164-6430, USA. HRP’s visit to Instituto Virologia Vegetale, CNR, Turin, Italy, was supported by a fellowship from the OECD under the Cooperative Research Program.

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Authors and Affiliations

  1. Department of Plant Pathology, Washington State University, PO Box 646430, Pullman, WA, 99164, USA

    Ying Zhai, Sudeep Bag & Hanu R. Pappu

  2. QAAFI, University of Queensland, St Lucia, QLD, Australia

    Neena Mitter

  3. Istituto di Virologia Vegetale, CNR, Strada delle Cacce 75, 10135, Turin, Italy

    Massimo Turina

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  1. Ying Zhai
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  2. Sudeep Bag
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Correspondence to Hanu R. Pappu.

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Zhai, Y., Bag, S., Mitter, N. et al. Mutational analysis of two highly conserved motifs in the silencing suppressor encoded by tomato spotted wilt virus (genus Tospovirus, family Bunyaviridae). Arch Virol 159, 1499–1504 (2014). https://doi.org/10.1007/s00705-013-1928-8

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