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Non-canonical targets destabilize microRNAs in human Argonautes

Although much is known about microRNA (miRNA) biogenesis and the mechanism by which miRNAs regulate their targets, little is known about the regulation of miRNA stability. Mature miRNAs are stabilized by binding to Argonaute (Ago) proteins, the core components of the RNA-induced silencing complex (R...

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Autores principales: Park, June Hyun, Shin, Sang-Yoon, Shin, Chanseok
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5389725/
https://www.ncbi.nlm.nih.gov/pubmed/28119422
http://dx.doi.org/10.1093/nar/gkx029
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author Park, June Hyun
Shin, Sang-Yoon
Shin, Chanseok
author_facet Park, June Hyun
Shin, Sang-Yoon
Shin, Chanseok
author_sort Park, June Hyun
collection PubMed
description Although much is known about microRNA (miRNA) biogenesis and the mechanism by which miRNAs regulate their targets, little is known about the regulation of miRNA stability. Mature miRNAs are stabilized by binding to Argonaute (Ago) proteins, the core components of the RNA-induced silencing complex (RISC). Recent studies suggest that interactions between miRNAs and their highly complementary target RNAs promote release of miRNAs from Ago proteins, and this in turn can lead to destabilization of miRNAs. However, the physiological triggers of miRNA destabilization with molecular mechanisms remain largely unknown. Here, using an in vitro system that consists of a minimal human Ago2–RISC in HEK293T cell lysates, we sought to understand how miRNAs are destabilized by their targets. Strikingly, we showed that miRNA destabilization is dramatically enhanced by an interaction with seedless, non-canonical targets. We then showed that this process entails not only unloading of miRNAs from Ago, but also 3΄ end destabilization of miRNAs occurred within Ago. Furthermore, our mutation analysis indicates that conformational changes in the hinge region of the Ago PAZ domain are likely to be the main driving force of the miRNA destabilization. Our collective results suggest that non-canonical targets may provide a stability control mechanism in the regulation of miRNAs in humans.
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spelling pubmed-53897252017-04-24 Non-canonical targets destabilize microRNAs in human Argonautes Park, June Hyun Shin, Sang-Yoon Shin, Chanseok Nucleic Acids Res NAR Breakthrough Article Although much is known about microRNA (miRNA) biogenesis and the mechanism by which miRNAs regulate their targets, little is known about the regulation of miRNA stability. Mature miRNAs are stabilized by binding to Argonaute (Ago) proteins, the core components of the RNA-induced silencing complex (RISC). Recent studies suggest that interactions between miRNAs and their highly complementary target RNAs promote release of miRNAs from Ago proteins, and this in turn can lead to destabilization of miRNAs. However, the physiological triggers of miRNA destabilization with molecular mechanisms remain largely unknown. Here, using an in vitro system that consists of a minimal human Ago2–RISC in HEK293T cell lysates, we sought to understand how miRNAs are destabilized by their targets. Strikingly, we showed that miRNA destabilization is dramatically enhanced by an interaction with seedless, non-canonical targets. We then showed that this process entails not only unloading of miRNAs from Ago, but also 3΄ end destabilization of miRNAs occurred within Ago. Furthermore, our mutation analysis indicates that conformational changes in the hinge region of the Ago PAZ domain are likely to be the main driving force of the miRNA destabilization. Our collective results suggest that non-canonical targets may provide a stability control mechanism in the regulation of miRNAs in humans. Oxford University Press 2017-02-28 2017-01-23 /pmc/articles/PMC5389725/ /pubmed/28119422 http://dx.doi.org/10.1093/nar/gkx029 Text en © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle NAR Breakthrough Article
Park, June Hyun
Shin, Sang-Yoon
Shin, Chanseok
Non-canonical targets destabilize microRNAs in human Argonautes
title Non-canonical targets destabilize microRNAs in human Argonautes
title_full Non-canonical targets destabilize microRNAs in human Argonautes
title_fullStr Non-canonical targets destabilize microRNAs in human Argonautes
title_full_unstemmed Non-canonical targets destabilize microRNAs in human Argonautes
title_short Non-canonical targets destabilize microRNAs in human Argonautes
title_sort non-canonical targets destabilize micrornas in human argonautes
topic NAR Breakthrough Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5389725/
https://www.ncbi.nlm.nih.gov/pubmed/28119422
http://dx.doi.org/10.1093/nar/gkx029
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