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Microprocessor dynamics shows co- and post-transcriptional processing of pri-miRNAs
miRNAs are small regulatory RNAs involved in the regulation of translation of target transcripts. miRNA biogenesis is a multistep process starting with the cleavage of the primary miRNA transcript in the nucleus by the Microprocessor complex. Endogenous processing of pri-miRNAs is challenging to stu...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cold Spring Harbor Laboratory Press
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5435862/ https://www.ncbi.nlm.nih.gov/pubmed/28250203 http://dx.doi.org/10.1261/rna.060715.117 |
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author | Louloupi, Annita Ntini, Evgenia Liz, Julia Ørom, Ulf Andersson |
author_facet | Louloupi, Annita Ntini, Evgenia Liz, Julia Ørom, Ulf Andersson |
author_sort | Louloupi, Annita |
collection | PubMed |
description | miRNAs are small regulatory RNAs involved in the regulation of translation of target transcripts. miRNA biogenesis is a multistep process starting with the cleavage of the primary miRNA transcript in the nucleus by the Microprocessor complex. Endogenous processing of pri-miRNAs is challenging to study and the in vivo kinetics of this process is not known. Here, we present a method for determining the processing kinetics of pri-miRNAs within intact cells over time, using a pulse-chase approach to label transcribed RNA during 15 min, and follow the processing within a 1-hour window after labeling with bromouridine. We show that pri-miRNAs exhibit different processing kinetics ranging from fast over intermediate to slow processing, and we provide evidence that pri-miRNA processing can occur both cotranscriptionally and post-transcriptionally. |
format | Online Article Text |
id | pubmed-5435862 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Cold Spring Harbor Laboratory Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-54358622017-06-02 Microprocessor dynamics shows co- and post-transcriptional processing of pri-miRNAs Louloupi, Annita Ntini, Evgenia Liz, Julia Ørom, Ulf Andersson RNA Article miRNAs are small regulatory RNAs involved in the regulation of translation of target transcripts. miRNA biogenesis is a multistep process starting with the cleavage of the primary miRNA transcript in the nucleus by the Microprocessor complex. Endogenous processing of pri-miRNAs is challenging to study and the in vivo kinetics of this process is not known. Here, we present a method for determining the processing kinetics of pri-miRNAs within intact cells over time, using a pulse-chase approach to label transcribed RNA during 15 min, and follow the processing within a 1-hour window after labeling with bromouridine. We show that pri-miRNAs exhibit different processing kinetics ranging from fast over intermediate to slow processing, and we provide evidence that pri-miRNA processing can occur both cotranscriptionally and post-transcriptionally. Cold Spring Harbor Laboratory Press 2017-06 /pmc/articles/PMC5435862/ /pubmed/28250203 http://dx.doi.org/10.1261/rna.060715.117 Text en © 2017 Louloupi et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society http://creativecommons.org/licenses/by-nc/4.0/ This article, published in RNA, is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/. |
spellingShingle | Article Louloupi, Annita Ntini, Evgenia Liz, Julia Ørom, Ulf Andersson Microprocessor dynamics shows co- and post-transcriptional processing of pri-miRNAs |
title | Microprocessor dynamics shows co- and post-transcriptional processing of pri-miRNAs |
title_full | Microprocessor dynamics shows co- and post-transcriptional processing of pri-miRNAs |
title_fullStr | Microprocessor dynamics shows co- and post-transcriptional processing of pri-miRNAs |
title_full_unstemmed | Microprocessor dynamics shows co- and post-transcriptional processing of pri-miRNAs |
title_short | Microprocessor dynamics shows co- and post-transcriptional processing of pri-miRNAs |
title_sort | microprocessor dynamics shows co- and post-transcriptional processing of pri-mirnas |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5435862/ https://www.ncbi.nlm.nih.gov/pubmed/28250203 http://dx.doi.org/10.1261/rna.060715.117 |
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