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RNA-Binding Protein MSI2 Binds to miR-301a-3p and Facilitates Its Distribution in Mitochondria of Endothelial Cells

Numerous miRNAs have been detected in mitochondria, which play important roles in many physiological and pathophysiological processes. However, the dynamic changes of miRNA distribution in mitochondria and their mechanisms in reactive oxygen species (ROS)-induced endothelial injury remain unclear. T...

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Autores principales: Guo, Qian Qian, Gao, Jing, Wang, Xiao Wei, Yin, Xian Lun, Zhang, Shu Cui, Li, Xue, Chi, Lian Li, Zhou, Xiao Ming, Wang, Zhe, Zhang, Qun Ye
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7859344/
https://www.ncbi.nlm.nih.gov/pubmed/33553241
http://dx.doi.org/10.3389/fmolb.2020.609828
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author Guo, Qian Qian
Gao, Jing
Wang, Xiao Wei
Yin, Xian Lun
Zhang, Shu Cui
Li, Xue
Chi, Lian Li
Zhou, Xiao Ming
Wang, Zhe
Zhang, Qun Ye
author_facet Guo, Qian Qian
Gao, Jing
Wang, Xiao Wei
Yin, Xian Lun
Zhang, Shu Cui
Li, Xue
Chi, Lian Li
Zhou, Xiao Ming
Wang, Zhe
Zhang, Qun Ye
author_sort Guo, Qian Qian
collection PubMed
description Numerous miRNAs have been detected in mitochondria, which play important roles in many physiological and pathophysiological processes. However, the dynamic changes of miRNA distribution in mitochondria and their mechanisms in reactive oxygen species (ROS)-induced endothelial injury remain unclear. Therefore, miRNA levels in whole cells and mitochondria of H(2)O(2)-treated endothelial cells were analyzed by small RNA sequencing in the present study. The results showed that H(2)O(2) significantly reduced the relative mitochondrial distribution of dozens of miRNAs in human umbilical vein endothelial cells (HUVECs). Among the high-abundance miRNAs, miR-301a-3p has the most significant changes in the redistribution between cytosol and mitochondria confirmed by absolute quantitative polymerase chain reaction (qPCR). To unravel the mechanism of miR-301a-3p distribution in mitochondria, RNA pull-down followed by label-free quantitative proteomic analysis was performed, and RNA-binding protein Musashi RNA binding protein 2 (MSI2) was found to specifically bind to miR-301a-3p. Western blotting and immunofluorescence colocalization assay showed that MSI2 was located in mitochondria of various cell types. H(2)O(2) significantly downregulated MSI2 expression in whole endothelial cells, promoted the distribution of MSI2 in cytosol and decreased its distribution in the mitochondria. Moreover, overexpression of MSI2 increased the mitochondrial distribution of miR-301a-3p, whereas inhibition of MSI2 decreased its distribution in mitochondria. Thus, MSI2 might be responsible for the distribution of miR-301a-3p between cytosol and mitochondria in endothelial cells. Our findings revealed for the first time that MSI2 was involved in the regulation of miRNA distribution in mitochondria and provided valuable insight into the mechanism of mitochondrial distribution of miRNAs.
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spelling pubmed-78593442021-02-05 RNA-Binding Protein MSI2 Binds to miR-301a-3p and Facilitates Its Distribution in Mitochondria of Endothelial Cells Guo, Qian Qian Gao, Jing Wang, Xiao Wei Yin, Xian Lun Zhang, Shu Cui Li, Xue Chi, Lian Li Zhou, Xiao Ming Wang, Zhe Zhang, Qun Ye Front Mol Biosci Molecular Biosciences Numerous miRNAs have been detected in mitochondria, which play important roles in many physiological and pathophysiological processes. However, the dynamic changes of miRNA distribution in mitochondria and their mechanisms in reactive oxygen species (ROS)-induced endothelial injury remain unclear. Therefore, miRNA levels in whole cells and mitochondria of H(2)O(2)-treated endothelial cells were analyzed by small RNA sequencing in the present study. The results showed that H(2)O(2) significantly reduced the relative mitochondrial distribution of dozens of miRNAs in human umbilical vein endothelial cells (HUVECs). Among the high-abundance miRNAs, miR-301a-3p has the most significant changes in the redistribution between cytosol and mitochondria confirmed by absolute quantitative polymerase chain reaction (qPCR). To unravel the mechanism of miR-301a-3p distribution in mitochondria, RNA pull-down followed by label-free quantitative proteomic analysis was performed, and RNA-binding protein Musashi RNA binding protein 2 (MSI2) was found to specifically bind to miR-301a-3p. Western blotting and immunofluorescence colocalization assay showed that MSI2 was located in mitochondria of various cell types. H(2)O(2) significantly downregulated MSI2 expression in whole endothelial cells, promoted the distribution of MSI2 in cytosol and decreased its distribution in the mitochondria. Moreover, overexpression of MSI2 increased the mitochondrial distribution of miR-301a-3p, whereas inhibition of MSI2 decreased its distribution in mitochondria. Thus, MSI2 might be responsible for the distribution of miR-301a-3p between cytosol and mitochondria in endothelial cells. Our findings revealed for the first time that MSI2 was involved in the regulation of miRNA distribution in mitochondria and provided valuable insight into the mechanism of mitochondrial distribution of miRNAs. Frontiers Media S.A. 2021-01-21 /pmc/articles/PMC7859344/ /pubmed/33553241 http://dx.doi.org/10.3389/fmolb.2020.609828 Text en Copyright © 2021 Guo, Gao, Wang, Yin, Zhang, Li, Chi, Zhou, Wang and Zhang. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Molecular Biosciences
Guo, Qian Qian
Gao, Jing
Wang, Xiao Wei
Yin, Xian Lun
Zhang, Shu Cui
Li, Xue
Chi, Lian Li
Zhou, Xiao Ming
Wang, Zhe
Zhang, Qun Ye
RNA-Binding Protein MSI2 Binds to miR-301a-3p and Facilitates Its Distribution in Mitochondria of Endothelial Cells
title RNA-Binding Protein MSI2 Binds to miR-301a-3p and Facilitates Its Distribution in Mitochondria of Endothelial Cells
title_full RNA-Binding Protein MSI2 Binds to miR-301a-3p and Facilitates Its Distribution in Mitochondria of Endothelial Cells
title_fullStr RNA-Binding Protein MSI2 Binds to miR-301a-3p and Facilitates Its Distribution in Mitochondria of Endothelial Cells
title_full_unstemmed RNA-Binding Protein MSI2 Binds to miR-301a-3p and Facilitates Its Distribution in Mitochondria of Endothelial Cells
title_short RNA-Binding Protein MSI2 Binds to miR-301a-3p and Facilitates Its Distribution in Mitochondria of Endothelial Cells
title_sort rna-binding protein msi2 binds to mir-301a-3p and facilitates its distribution in mitochondria of endothelial cells
topic Molecular Biosciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7859344/
https://www.ncbi.nlm.nih.gov/pubmed/33553241
http://dx.doi.org/10.3389/fmolb.2020.609828
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