Pre-microRNA and mature microRNA in human mitochondria.

<h4>Background</h4>Because of the central functions of the mitochondria in providing metabolic energy and initiating apoptosis on one hand and the role that microRNA (miRNA) play in gene expression, we hypothesized that some miRNA could be present in the mitochondria for post-transcripto...

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Autores principales:	Eric Barrey, Gaelle Saint-Auret, Blandine Bonnamy, Dominique Damas, Orane Boyer, Xavier Gidrol
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2011
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/6055ed26194d467487f535d0152950c3
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spelling	oai:doaj.org-article:6055ed26194d467487f535d0152950c32021-11-18T06:53:08ZPre-microRNA and mature microRNA in human mitochondria.1932-620310.1371/journal.pone.0020220https://doaj.org/article/6055ed26194d467487f535d0152950c32011-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21637849/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>Because of the central functions of the mitochondria in providing metabolic energy and initiating apoptosis on one hand and the role that microRNA (miRNA) play in gene expression, we hypothesized that some miRNA could be present in the mitochondria for post-transcriptomic regulation by RNA interference. We intend to identify miRNA localized in the mitochondria isolated from human skeletal primary muscular cells.<h4>Methodology/principal findings</h4>To investigate the potential origin of mitochondrial miRNA, we in-silico searched for microRNA candidates in the mtDNA. Twenty five human pre-miRNA and 33 miRNA aligments (E-value<0.1) were found in the reference mitochondrial sequence and some of the best candidates were chosen for a co-localization test. In situ hybridization of pre-mir-302a, pre-let-7b and mir-365, using specific labelled locked nucleic acids and confocal microscopy, demonstrated that these miRNA were localized in mitochondria of human myoblasts. Total RNA was extracted from enriched mitochondria isolated by an immunomagnetic method from a culture of human myotubes. The detection of 742 human miRNA (miRBase) were monitored by RT-qPCR at three increasing mtRNA inputs. Forty six miRNA were significantly expressed (2(nd) derivative method Cp>35) for the smallest RNA input concentration and 204 miRNA for the maximum RNA input concentration. In silico analysis predicted 80 putative miRNA target sites in the mitochondrial genome (E-value<0.05).<h4>Conclusions/significance</h4>The present study experimentally demonstrated for the first time the presence of pre-miRNA and miRNA in the human mitochondria isolated from skeletal muscular cells. A set of miRNA were significantly detected in mitochondria fraction. The origin of these pre-miRNA and miRNA should be further investigate to determine if they are imported from the cytosol and/or if they are partially processed in the mitochondria.Eric BarreyGaelle Saint-AuretBlandine BonnamyDominique DamasOrane BoyerXavier GidrolPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 6, Iss 5, p e20220 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Eric Barrey Gaelle Saint-Auret Blandine Bonnamy Dominique Damas Orane Boyer Xavier Gidrol Pre-microRNA and mature microRNA in human mitochondria.
description	<h4>Background</h4>Because of the central functions of the mitochondria in providing metabolic energy and initiating apoptosis on one hand and the role that microRNA (miRNA) play in gene expression, we hypothesized that some miRNA could be present in the mitochondria for post-transcriptomic regulation by RNA interference. We intend to identify miRNA localized in the mitochondria isolated from human skeletal primary muscular cells.<h4>Methodology/principal findings</h4>To investigate the potential origin of mitochondrial miRNA, we in-silico searched for microRNA candidates in the mtDNA. Twenty five human pre-miRNA and 33 miRNA aligments (E-value<0.1) were found in the reference mitochondrial sequence and some of the best candidates were chosen for a co-localization test. In situ hybridization of pre-mir-302a, pre-let-7b and mir-365, using specific labelled locked nucleic acids and confocal microscopy, demonstrated that these miRNA were localized in mitochondria of human myoblasts. Total RNA was extracted from enriched mitochondria isolated by an immunomagnetic method from a culture of human myotubes. The detection of 742 human miRNA (miRBase) were monitored by RT-qPCR at three increasing mtRNA inputs. Forty six miRNA were significantly expressed (2(nd) derivative method Cp>35) for the smallest RNA input concentration and 204 miRNA for the maximum RNA input concentration. In silico analysis predicted 80 putative miRNA target sites in the mitochondrial genome (E-value<0.05).<h4>Conclusions/significance</h4>The present study experimentally demonstrated for the first time the presence of pre-miRNA and miRNA in the human mitochondria isolated from skeletal muscular cells. A set of miRNA were significantly detected in mitochondria fraction. The origin of these pre-miRNA and miRNA should be further investigate to determine if they are imported from the cytosol and/or if they are partially processed in the mitochondria.
format	article
author	Eric Barrey Gaelle Saint-Auret Blandine Bonnamy Dominique Damas Orane Boyer Xavier Gidrol
author_facet	Eric Barrey Gaelle Saint-Auret Blandine Bonnamy Dominique Damas Orane Boyer Xavier Gidrol
author_sort	Eric Barrey
title	Pre-microRNA and mature microRNA in human mitochondria.
title_short	Pre-microRNA and mature microRNA in human mitochondria.
title_full	Pre-microRNA and mature microRNA in human mitochondria.
title_fullStr	Pre-microRNA and mature microRNA in human mitochondria.
title_full_unstemmed	Pre-microRNA and mature microRNA in human mitochondria.
title_sort	pre-microrna and mature microrna in human mitochondria.
publisher	Public Library of Science (PLoS)
publishDate	2011
url	https://doaj.org/article/6055ed26194d467487f535d0152950c3
work_keys_str_mv	AT ericbarrey premicrornaandmaturemicrornainhumanmitochondria AT gaellesaintauret premicrornaandmaturemicrornainhumanmitochondria AT blandinebonnamy premicrornaandmaturemicrornainhumanmitochondria AT dominiquedamas premicrornaandmaturemicrornainhumanmitochondria AT oraneboyer premicrornaandmaturemicrornainhumanmitochondria AT xaviergidrol premicrornaandmaturemicrornainhumanmitochondria
_version_	1718424232544698368

Pre-microRNA and mature microRNA in human mitochondria.

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