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Functional analysis of three Arabidopsis ARGONAUTES using slicer-defective mutants

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Functional analysis of three Arabidopsis ARGONAUTES using slicer-defective mutants

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dc.contributor.author CARBONELL, ALBERTO es_ES
dc.contributor.author Fahlgren, Noah es_ES
dc.contributor.author García-Ruíz, Hernan es_ES
dc.contributor.author Gilbert, Kerrigan B. es_ES
dc.contributor.author Montgomery, Taiowa A. es_ES
dc.contributor.author Nguyen, Tammy es_ES
dc.contributor.author Cuperus, Josh T. es_ES
dc.contributor.author Carrington, James C. es_ES
dc.date.accessioned 2021-02-10T04:31:25Z
dc.date.available 2021-02-10T04:31:25Z
dc.date.issued 2012-09 es_ES
dc.identifier.issn 1040-4651 es_ES
dc.identifier.uri http://hdl.handle.net/10251/160977
dc.description.abstract [EN] In RNA-directed silencing pathways, ternary complexes result from small RNA-guided ARGONAUTE (AGO) associating with target transcripts. Target transcripts are often silenced through direct cleavage (slicing), destabilization through slicer-independent turnover mechanisms, and translational repression. Here, wild-type and active-site defective forms of several Arabidopsis thaliana AGO proteins involved in posttranscriptional silencing were used to examine several AGO functions, including small RNA binding, interaction with target RNA, slicing or destabilization of target RNA, secondary small interfering RNA formation, and antiviral activity. Complementation analyses in ago mutant plants revealed that the catalytic residues of AGO1, AGO2, and AGO7 are required to restore the defects of Arabidopsis ago1-25, ago2-1, and zip-1 (AGO7-defective) mutants, respectively. AGO2 had slicer activity in transient assays but could not trigger secondary small interfering RNA biogenesis, and catalytically active AGO2 was necessary for local and systemic antiviral activity against Turnip mosaic virus. Slicer-defective AGOs associated with miRNAs and stabilized AGO-miRNA-target RNA ternary complexes in individual target coimmunoprecipitation assays. In genome-wide AGO-miRNA-target RNA coimmunoprecipitation experiments, slicer-defective AGO1-miRNA associated with target RNA more effectively than did wild-type AGO1-miRNA. These data not only reveal functional roles for AGO1, AGO2, and AGO7 slicer activity, but also indicate an approach to capture ternary complexes more efficiently for genome-wide analyses. es_ES
dc.description.sponsorship We thank Goretti Nguyen for excellent technical assistance. A. C. was supported by a postdoctoral fellowship from the Ministerio de Ciencia e Innovacion (BMC-2008-0188). H.G.-R. was the recipient of a Helen Hay Whitney Postdoctoral fellowship (F-972). This work was supported by grants from the National Science Foundation (MCB-1231726), the National Institutes of Health (AI043288), and Monsanto Corporation. es_ES
dc.language Inglés es_ES
dc.publisher American Society of Plant Biologists es_ES
dc.relation.ispartof The Plant Cell es_ES
dc.rights Reserva de todos los derechos es_ES
dc.subject MiRNA es_ES
dc.subject Argonaute es_ES
dc.subject RNA silencing es_ES
dc.title Functional analysis of three Arabidopsis ARGONAUTES using slicer-defective mutants es_ES
dc.type Artículo es_ES
dc.identifier.doi 10.1105/tpc.112.099945 es_ES
dc.relation.projectID info:eu-repo/grantAgreement/NSF//1231726/US/Function of Arabidopsis Small RNA-ARGONAUTE Complexes/ es_ES
dc.relation.projectID info:eu-repo/grantAgreement/NIH//AI043288/ es_ES
dc.relation.projectID info:eu-repo/grantAgreement/MICINN//BMC-2008-0188/ es_ES
dc.relation.projectID info:eu-repo/grantAgreement/HHWF//F-972/ 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 Carbonell, A.; Fahlgren, N.; García-Ruíz, H.; Gilbert, KB.; Montgomery, TA.; Nguyen, T.; Cuperus, JT.... (2012). Functional analysis of three Arabidopsis ARGONAUTES using slicer-defective mutants. The Plant Cell. 24(9):3613-3629. https://doi.org/10.1105/tpc.112.099945 es_ES
dc.description.accrualMethod S es_ES
dc.relation.publisherversion https://doi.org/10.1105/tpc.112.099945 es_ES
dc.description.upvformatpinicio 3613 es_ES
dc.description.upvformatpfin 3629 es_ES
dc.type.version info:eu-repo/semantics/publishedVersion es_ES
dc.description.volume 24 es_ES
dc.description.issue 9 es_ES
dc.identifier.pmid 23023169 es_ES
dc.identifier.pmcid PMC3480291 es_ES
dc.relation.pasarela S\377810 es_ES
dc.contributor.funder Helen Hay Whitney Foundation es_ES
dc.contributor.funder National Science Foundation, EEUU es_ES
dc.contributor.funder Ministerio de Ciencia e Innovación es_ES
dc.contributor.funder National Institutes of Health, EEUU es_ES
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