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Actin-binding protein regulation by microRNAs as a novel microbial strategy to modulate phagocytosis by host cells: the case of N-Wasp and miR-142-3p
Mycobacterium tuberculosis (Mtb) is a successful intracellular pathogen that thrives in macrophages (Mφs). There is a need to better understand how Mtb alters cellular processes like phagolysosome biogenesis, a classical determinant of its pathogenesis. A central feature of this bacteria's stra...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3672780/ https://www.ncbi.nlm.nih.gov/pubmed/23760605 http://dx.doi.org/10.3389/fcimb.2013.00019 |
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author | Bettencourt, Paulo Marion, Sabrina Pires, David Santos, Leonor F. Lastrucci, Claire Carmo, Nuno Blake, Jonathon Benes, Vladimir Griffiths, Gareth Neyrolles, Olivier Lugo-Villarino, Geanncarlo Anes, Elsa |
author_facet | Bettencourt, Paulo Marion, Sabrina Pires, David Santos, Leonor F. Lastrucci, Claire Carmo, Nuno Blake, Jonathon Benes, Vladimir Griffiths, Gareth Neyrolles, Olivier Lugo-Villarino, Geanncarlo Anes, Elsa |
author_sort | Bettencourt, Paulo |
collection | PubMed |
description | Mycobacterium tuberculosis (Mtb) is a successful intracellular pathogen that thrives in macrophages (Mφs). There is a need to better understand how Mtb alters cellular processes like phagolysosome biogenesis, a classical determinant of its pathogenesis. A central feature of this bacteria's strategy is the manipulation of Mφ actin. Here, we examined the role of microRNAs (miRNAs) as a potential mechanism in the regulation of actin-mediated events leading to phagocytosis in the context of mycobacteria infection. Given that non-virulent Mycobacterium smegmatis also controls actin filament assembly to prolong its intracellular survival inside host cells, we performed a global transcriptomic analysis to assess the modulation of miRNAs upon M. smegmatis infection of the murine Mφ cell line, J774A.1. This approach identified miR-142-3p as a key candidate to be involved in the regulation of actin dynamics required in phagocytosis. We unequivocally demonstrate that miR-142-3p targets N-Wasp, an actin-binding protein required during microbial challenge. A gain-of-function approach for miR-142-3p revealed a down-regulation of N-Wasp expression accompanied by a decrease of mycobacteria intake, while a loss-of-function approach yielded the reciprocal increase of the phagocytosis process. Equally important, we show Mtb induces the early expression of miR-142-3p and partially down-regulates N-Wasp protein levels in both the murine J774A.1 cell line and primary human Mφs. As proof of principle, the partial siRNA-mediated knock down of N-Wasp resulted in a decrease of Mtb intake by human Mφs, reflected in lower levels of colony-forming units (CFU) counts over time. We therefore propose the modulation of miRNAs as a novel strategy in mycobacterial infection to control factors involved in actin filament assembly and other early events of phagolysosome biogenesis. |
format | Online Article Text |
id | pubmed-3672780 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-36727802013-06-11 Actin-binding protein regulation by microRNAs as a novel microbial strategy to modulate phagocytosis by host cells: the case of N-Wasp and miR-142-3p Bettencourt, Paulo Marion, Sabrina Pires, David Santos, Leonor F. Lastrucci, Claire Carmo, Nuno Blake, Jonathon Benes, Vladimir Griffiths, Gareth Neyrolles, Olivier Lugo-Villarino, Geanncarlo Anes, Elsa Front Cell Infect Microbiol Microbiology Mycobacterium tuberculosis (Mtb) is a successful intracellular pathogen that thrives in macrophages (Mφs). There is a need to better understand how Mtb alters cellular processes like phagolysosome biogenesis, a classical determinant of its pathogenesis. A central feature of this bacteria's strategy is the manipulation of Mφ actin. Here, we examined the role of microRNAs (miRNAs) as a potential mechanism in the regulation of actin-mediated events leading to phagocytosis in the context of mycobacteria infection. Given that non-virulent Mycobacterium smegmatis also controls actin filament assembly to prolong its intracellular survival inside host cells, we performed a global transcriptomic analysis to assess the modulation of miRNAs upon M. smegmatis infection of the murine Mφ cell line, J774A.1. This approach identified miR-142-3p as a key candidate to be involved in the regulation of actin dynamics required in phagocytosis. We unequivocally demonstrate that miR-142-3p targets N-Wasp, an actin-binding protein required during microbial challenge. A gain-of-function approach for miR-142-3p revealed a down-regulation of N-Wasp expression accompanied by a decrease of mycobacteria intake, while a loss-of-function approach yielded the reciprocal increase of the phagocytosis process. Equally important, we show Mtb induces the early expression of miR-142-3p and partially down-regulates N-Wasp protein levels in both the murine J774A.1 cell line and primary human Mφs. As proof of principle, the partial siRNA-mediated knock down of N-Wasp resulted in a decrease of Mtb intake by human Mφs, reflected in lower levels of colony-forming units (CFU) counts over time. We therefore propose the modulation of miRNAs as a novel strategy in mycobacterial infection to control factors involved in actin filament assembly and other early events of phagolysosome biogenesis. Frontiers Media S.A. 2013-06-05 /pmc/articles/PMC3672780/ /pubmed/23760605 http://dx.doi.org/10.3389/fcimb.2013.00019 Text en Copyright © 2013 Bettencourt, Marion, Pires, Santos, Lastrucci, Carmo, Blake, Benes, Griffiths, Neyrolles, Lugo-Villarino and Anes. http://creativecommons.org/licenses/by/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in other forums, provided the original authors and source are credited and subject to any copyright notices concerning any third-party graphics etc. |
spellingShingle | Microbiology Bettencourt, Paulo Marion, Sabrina Pires, David Santos, Leonor F. Lastrucci, Claire Carmo, Nuno Blake, Jonathon Benes, Vladimir Griffiths, Gareth Neyrolles, Olivier Lugo-Villarino, Geanncarlo Anes, Elsa Actin-binding protein regulation by microRNAs as a novel microbial strategy to modulate phagocytosis by host cells: the case of N-Wasp and miR-142-3p |
title | Actin-binding protein regulation by microRNAs as a novel microbial strategy to modulate phagocytosis by host cells: the case of N-Wasp and miR-142-3p |
title_full | Actin-binding protein regulation by microRNAs as a novel microbial strategy to modulate phagocytosis by host cells: the case of N-Wasp and miR-142-3p |
title_fullStr | Actin-binding protein regulation by microRNAs as a novel microbial strategy to modulate phagocytosis by host cells: the case of N-Wasp and miR-142-3p |
title_full_unstemmed | Actin-binding protein regulation by microRNAs as a novel microbial strategy to modulate phagocytosis by host cells: the case of N-Wasp and miR-142-3p |
title_short | Actin-binding protein regulation by microRNAs as a novel microbial strategy to modulate phagocytosis by host cells: the case of N-Wasp and miR-142-3p |
title_sort | actin-binding protein regulation by micrornas as a novel microbial strategy to modulate phagocytosis by host cells: the case of n-wasp and mir-142-3p |
topic | Microbiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3672780/ https://www.ncbi.nlm.nih.gov/pubmed/23760605 http://dx.doi.org/10.3389/fcimb.2013.00019 |
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