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Functional dissection of a plant Argonaute

RNA guided ribonuclease complexes play central role in RNA interference. Members of the evolutionarily conserved Argonaute protein family form the catalytic cores of these complexes. Unlike a number of other plant Argonautes, the role of AGO2 has been obscure until recently. Newer data, however, hav...

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Autores principales: Fátyol, Károly, Ludman, Márta, Burgyán, József
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2016
Materias:
RNA
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4756824/
https://www.ncbi.nlm.nih.gov/pubmed/26673719
http://dx.doi.org/10.1093/nar/gkv1371
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author Fátyol, Károly
Ludman, Márta
Burgyán, József
author_facet Fátyol, Károly
Ludman, Márta
Burgyán, József
author_sort Fátyol, Károly
collection PubMed
description RNA guided ribonuclease complexes play central role in RNA interference. Members of the evolutionarily conserved Argonaute protein family form the catalytic cores of these complexes. Unlike a number of other plant Argonautes, the role of AGO2 has been obscure until recently. Newer data, however, have indicated its involvement in various biotic and abiotic stress responses. Despite its suggested importance, there is no detailed characterization of this protein to date. Here we report cloning and molecular characterization of the AGO2 protein of the virological model plant Nicotiana benthamiana. We show that AGO2 can directly repress translation via various miRNA target site constellations (ORF, 3′ UTR). Interestingly, although AGO2 seems to be able to silence gene expression in a slicing independent fashion, its catalytic activity is still a prerequisite for efficient translational repression. Additionally, mismatches between the 3′ end of the miRNA guide strand and the 5′ end of the target site enhance gene silencing by AGO2. Several functionally important amino acid residues of AGO2 have been identified that affect its small RNA loading, cleavage activity, translational repression potential and antiviral activity. The data presented here help us to understand how AGO2 aids plants to deal with stress.
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spelling pubmed-47568242016-02-18 Functional dissection of a plant Argonaute Fátyol, Károly Ludman, Márta Burgyán, József Nucleic Acids Res RNA RNA guided ribonuclease complexes play central role in RNA interference. Members of the evolutionarily conserved Argonaute protein family form the catalytic cores of these complexes. Unlike a number of other plant Argonautes, the role of AGO2 has been obscure until recently. Newer data, however, have indicated its involvement in various biotic and abiotic stress responses. Despite its suggested importance, there is no detailed characterization of this protein to date. Here we report cloning and molecular characterization of the AGO2 protein of the virological model plant Nicotiana benthamiana. We show that AGO2 can directly repress translation via various miRNA target site constellations (ORF, 3′ UTR). Interestingly, although AGO2 seems to be able to silence gene expression in a slicing independent fashion, its catalytic activity is still a prerequisite for efficient translational repression. Additionally, mismatches between the 3′ end of the miRNA guide strand and the 5′ end of the target site enhance gene silencing by AGO2. Several functionally important amino acid residues of AGO2 have been identified that affect its small RNA loading, cleavage activity, translational repression potential and antiviral activity. The data presented here help us to understand how AGO2 aids plants to deal with stress. Oxford University Press 2016-02-18 2015-12-15 /pmc/articles/PMC4756824/ /pubmed/26673719 http://dx.doi.org/10.1093/nar/gkv1371 Text en © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle RNA
Fátyol, Károly
Ludman, Márta
Burgyán, József
Functional dissection of a plant Argonaute
title Functional dissection of a plant Argonaute
title_full Functional dissection of a plant Argonaute
title_fullStr Functional dissection of a plant Argonaute
title_full_unstemmed Functional dissection of a plant Argonaute
title_short Functional dissection of a plant Argonaute
title_sort functional dissection of a plant argonaute
topic RNA
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4756824/
https://www.ncbi.nlm.nih.gov/pubmed/26673719
http://dx.doi.org/10.1093/nar/gkv1371
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