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Key importance of small RNA binding for the activity of a GW motif-containing RNA silencing suppressor

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Key importance of small RNA binding for the activity of a GW motif-containing RNA silencing suppressor

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dc.contributor.author Perez-Cañamas, M es_ES
dc.contributor.author Hernandez Fort, Carmen es_ES
dc.date.accessioned 2016-07-25T11:36:21Z
dc.date.available 2016-07-25T11:36:21Z
dc.date.issued 2015-01-30
dc.identifier.issn 0021-9258
dc.identifier.uri http://hdl.handle.net/10251/68095
dc.description.abstract Background: GW/WG motif-containing viral suppressors of RNA silencing (VSRs) have been proposed to act through interaction with Argonaute (AGO) proteins. Results: The activity of a GW motif-containing VSR was found to rely on small RNA binding capability rather than on AGO interaction. Conclusion: Overlapping signals in VSRs may lead to misinterpretation of relevant molecular traits. Significance: Knowing primary target(s) of VSRs is critical for better understanding of the host-virus arms race. Viruses express viral suppressors of RNA silencing (VSRs) to counteract RNA silencing-based host defenses. Although virtually all stages of the antiviral silencing pathway can be inhibited by VSRs, small RNAs (sRNAs) and Argonaute (AGO) proteins seem to be the most frequent targets. Recently, GW/WG motifs of some VSRs have been proposed to dictate their suppressor function by mediating interaction with AGO(s). Here we have studied the VSR encoded by Pelargonium line pattern virus (family Tombusviridae). The results show that p37, the viral coat protein, blocks RNA silencing. Site-directed mutagenesis of some p37 sequence traits, including a conserved GW motif, allowed generation of suppressor-competent and -incompetent molecules and uncoupling of the VSR and particle assembly capacities. The engineered mutants were used to assess the importance of p37 functions for viral infection and the relative contribution of diverse molecular interactions to suppressor activity. Two main conclusions can be drawn: (i) the silencing suppression and encapsidation functions of p37 are both required for systemic Pelargonium line pattern virus infection, and (ii) the suppressor activity of p37 relies on the ability to bind sRNAs rather than on interaction with AGOs. The data also caution against potential misinterpretations of results due to overlap of sequence signals related to distinct protein properties. This is well illustrated by mutation of the GW motif in p37 that concurrently affects nucleolar localization, efficient interaction with AGO1, and sRNA binding capability. These concomitant effects could have been overlooked in other GW motif-containing suppressors, as we exemplify with the orthologous p38 of turnip crinkle virus. es_ES
dc.description.sponsorship This work was supported by Grant BFU2012-36095 from the Ministerio de Economia y Competitividad (MINECO, Spain) ( to C. H.). en_EN
dc.language Inglés es_ES
dc.publisher American Society for Biochemistry and Molecular Biology es_ES
dc.relation.ispartof Journal of Biological Chemistry es_ES
dc.rights Reserva de todos los derechos es_ES
dc.subject Argonaute es_ES
dc.subject Host-Pathogen Interaction es_ES
dc.subject Nucleolus es_ES
dc.subject Plant Virus es_ES
dc.subject RNA Virus es_ES
dc.subject GW es_ES
dc.subject WG Motifs es_ES
dc.subject Pelargonium Line Pattern Virus es_ES
dc.subject Coat Protein es_ES
dc.subject sRNA Binding es_ES
dc.subject Viral Suppressor of RNA Silencing es_ES
dc.title Key importance of small RNA binding for the activity of a GW motif-containing RNA silencing suppressor es_ES
dc.type Artículo es_ES
dc.identifier.doi 10.1074/jbc.M114.593707
dc.relation.projectID info:eu-repo/grantAgreement/MINECO//BFU2012-36095/ES/ANALISIS DE UNA RELACION COMENSALISTA VIRUS-PLANTA: ESTUDIO DE DETERMINANTES DE ACUMULACION VIRAL Y DE POSIBLES ALTERACIONES EPIGENETICAS EN EL GENOMA DEL HUESPED/ es_ES
dc.rights.accessRights Abierto es_ES
dc.contributor.affiliation Universitat Politècnica de València. Instituto Universitario Mixto de Biología Molecular y Celular de Plantas - Institut Universitari Mixt de Biologia Molecular i Cel·lular de Plantes es_ES
dc.description.bibliographicCitation Perez-Cañamas, M.; Hernandez Fort, C. (2015). Key importance of small RNA binding for the activity of a GW motif-containing RNA silencing suppressor. Journal of Biological Chemistry. 290(5):3106-3120. https://doi.org/10.1074/jbc.M114.593707 es_ES
dc.description.accrualMethod S es_ES
dc.relation.publisherversion http://dx.doi.org/10.1074/jbc.M114.593707 es_ES
dc.description.upvformatpinicio 3106 es_ES
dc.description.upvformatpfin 3120 es_ES
dc.type.version info:eu-repo/semantics/publishedVersion es_ES
dc.description.volume 290 es_ES
dc.description.issue 5 es_ES
dc.relation.senia 305708 es_ES
dc.identifier.pmcid PMC4317005 en_EN
dc.contributor.funder Ministerio de Economía y Competitividad es_ES
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