Functional analysis of three Arabidopsis ARGONAUTES using slicer-defective mutants

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dc.contributor.affiliationInstituto Universitario Mixto de Biología Molecular y Celular de Plantas
dc.contributor.authorCARBONELL, ALBERTO
dc.contributor.authorFahlgren, Noahes_ES
dc.contributor.authorGarcía-Ruíz, Hernanes_ES
dc.contributor.authorGilbert, Kerrigan B.es_ES
dc.contributor.authorMontgomery, Taiowa A.es_ES
dc.contributor.authorNguyen, Tammyes_ES
dc.contributor.authorCuperus, Josh T.es_ES
dc.contributor.authorCarrington, James C.es_ES
dc.contributor.funderHelen Hay Whitney Foundationes_ES
dc.contributor.funderNational Science Foundation, EEUUes_ES
dc.contributor.funderMinisterio de Ciencia e Innovaciónes_ES
dc.contributor.funderNational Institutes of Health, EEUUes_ES
dc.date.accessioned2021-02-10T04:31:25Z
dc.date.available2021-02-10T04:31:25Z
dc.date.issued2012-09es_ES
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.en_EN
dc.description.accrualMethodSes_ES
dc.description.bibliographicCitationCarbonell, 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.112.099945es_ES
dc.description.issue9es_ES
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dc.description.sponsorshipWe 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
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dc.identifier.doi10.1105/tpc.112.099945es_ES
dc.identifier.issn1040-4651es_ES
dc.identifier.pmcidPMC3480291es_ES
dc.identifier.pmid23023169es_ES
dc.identifier.urihttps://riunet.upv.es/handle/10251/160977
dc.languageIngléses_ES
dc.publisherAmerican Society of Plant Biologistses_ES
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dc.relation.projectIDinfo:eu-repo/grantAgreement/NSF//1231726/US/Function of Arabidopsis Small RNA-ARGONAUTE Complexes/es_ES
dc.relation.projectIDinfo:eu-repo/grantAgreement/NIH//AI043288/es_ES
dc.relation.projectIDinfo:eu-repo/grantAgreement/MICINN//BMC-2008-0188/es_ES
dc.relation.projectIDinfo:eu-repo/grantAgreement/HHWF//F-972/es_ES
dc.relation.publisherversionhttps://doi.org/10.1105/tpc.112.099945es_ES
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dc.rightsReserva de todos los derechoses_ES
dc.rights.accessRightsAbiertoes_ES
dc.subjectMiRNAes_ES
dc.subjectArgonautees_ES
dc.subjectRNA silencinges_ES
dc.titleFunctional analysis of three Arabidopsis ARGONAUTES using slicer-defective mutantses_ES
dc.typeArtículoes_ES
dc.type.versioninfo:eu-repo/semantics/publishedVersiones_ES
dspace.entity.typePublication
person.identifier46496
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CARBONELL;Fahlgren;García-Ruíz - Functional analysis of three Arabidopsis ARGONAUTES using slicer....pdf
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