NDLI: Tobacco rattle virus 16K silencing suppressor binds ARGONAUTE 4 and inhibits formation of RNA silencing complexes.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Fernández-Calvino, Lourdes Martínez-Priego, Llúcia Szabo, Edit Z. Guzmán-Benito, Irene González, Inmaculada Canto, Tomás Lakatos, Lóránt Llave, César
Description	Country affiliation: Spain Author Affiliation: Fernández-Calvino L ( 1âDepartment of Environmental Biology, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Ramiro de Maeztu 9, 28040 Madrid, Spain.); Martínez-Priego L ( 1âDepartment of Environmental Biology, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Ramiro de Maeztu 9, 28040 Madrid, Spain.); Szabo EZ ( 2âDepartment of Dermatology and Allergology, University of Szeged, H-6720 Szeged, Koranyi str. 6, Hungary.); Guzmán-Benito I ( 1âDepartment of Environmental Biology, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Ramiro de Maeztu 9, 28040 Madrid, Spain.); González I ( 1âDepartment of Environmental Biology, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Ramiro de Maeztu 9, 28040 Madrid, Spain.); Canto T ( 1âDepartment of Environmental Biology, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Ramiro de Maeztu 9, 28040 Madrid, Spain.); Lakatos L ( 2âDepartment of Dermatology and Allergology, University of Szeged, H-6720 Szeged, Koranyi str. 6, Hungary 3âMTA-SZTE Dermatological Research Group, Hungary.); Llave C ( 1âDepartment of Environmental Biology, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Ramiro de Maeztu 9, 28040 Madrid, Spain.)
Abstract	The cysteine-rich 16K protein of tobacco rattle virus (TRV), the type member of the genus Tobravirus, is known to suppress RNA silencing. However, the mechanism of action of the 16K suppressor is not well understood. In this study, we used a GFP-based sensor strategy and an Agrobacterium-mediated transient assay in Nicotiana benthamiana to show that 16K was unable to inhibit the activity of existing small interfering RNA (siRNA)- and microRNA (miRNA)-programmed RNA-induced silencing effector complexes (RISCs). In contrast, 16K efficiently interfered with de novo formation of miRNA- and siRNA-guided RISCs, thus preventing cleavage of target RNA. Interestingly, we found that transiently expressed endogenous miR399 and miR172 directed sequence-specific silencing of complementary sequences of viral origin. 16K failed to bind small RNAs, although it interacted with ARGONAUTE 4, as revealed by bimolecular fluorescence complementation and immunoprecipitation assays. Site-directed mutagenesis demonstrated that highly conserved cysteine residues within the N-terminal and central regions of the 16K protein are required for protein stability and/or RNA silencing suppression.
File Format	HTM / HTML
ISSN	00221317
Issue Number	1
Volume Number	97
e-ISSN	14652099
Journal	Journal of General Virology
Language	English
Publisher	Microbiology Society
Publisher Date	2016-01-01
Publisher Place	Great Britain (UK)
Access Restriction	Subscribed
Subject Keyword	Research Support, Non-u.s. Gov't Discipline Virology Viral Proteins Metabolism Rna Interference Journal Article Rna-binding Proteins Virology Plant Viruses Tobacco Host-pathogen Interactions Immunology Rna Viruses Protein Binding Physiology
Content Type	Text
Resource Type	Article
Subject	Virology

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