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 hijacks cellular processes like phagolysosome biogenesis, a classical pathogenesis determinant. A central feature to this microbial strategy is the...
| Main Authors: | , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2013-06-01
|
| Series: | Frontiers in Cellular and Infection Microbiology |
| Subjects: | |
| Online Access: | http://journal.frontiersin.org/Journal/10.3389/fcimb.2013.00019/full |
| _version_ | 1828808792359305216 |
|---|---|
| author | Paulo eBettencourt Sabrina eMarion David ePires Leonor eSantos Claire eLastrucci Nuno eCarmo Jonathon eBlake Vladimir eBenes Gareth eGriffiths Olivier eNeyrolles Geanncarlo eLugo-Villarino Elsa eAnes |
| author_facet | Paulo eBettencourt Sabrina eMarion David ePires Leonor eSantos Claire eLastrucci Nuno eCarmo Jonathon eBlake Vladimir eBenes Gareth eGriffiths Olivier eNeyrolles Geanncarlo eLugo-Villarino Elsa eAnes |
| author_sort | Paulo eBettencourt |
| collection | DOAJ |
| description | Mycobacterium tuberculosis (Mtb) is a successful intracellular pathogen that thrives in macrophages (Mφs). There is a need to better understand how Mtb hijacks cellular processes like phagolysosome biogenesis, a classical pathogenesis determinant. A central feature to this microbial strategy is the manipulation of actin in Mφs. Here, we examine 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 filamentation to prolong its instracellular survival inside host cells, our approach was to perform a global transcriptomic approach to assess the modulation of miRNAs upon M. smegmatis infection of the murine Mφ cell line, J774A.1. This approach yielded miR-142-3p as a top candidate to be involved in the regulation of actin dynamics required in phagocytosis. We unequivocally demonstrate 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 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 counts over time. We therefore propose the modulation of miRNAs as a novel strategy in mycobacterial infection to control factors involved in actin filamentation and other early events of phagolysosome biogenesis, such as the case presented here for miR-142-3p influencing N-Wasp activity during the phagocytosis process. |
| first_indexed | 2024-12-12T08:46:20Z |
| format | Article |
| id | doaj.art-ca535b5b47334e7b86b08543cab4d56e |
| institution | Directory Open Access Journal |
| issn | 2235-2988 |
| language | English |
| last_indexed | 2024-12-12T08:46:20Z |
| publishDate | 2013-06-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cellular and Infection Microbiology |
| spelling | doaj.art-ca535b5b47334e7b86b08543cab4d56e2022-12-22T00:30:31ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882013-06-01310.3389/fcimb.2013.0001950498Actin-binding protein regulation by microRNAs as a novel microbial strategy to modulate phagocytosis by host cells: the case of N-Wasp and miR-142-3pPaulo eBettencourt0Sabrina eMarion1David ePires2Leonor eSantos3Claire eLastrucci4Nuno eCarmo5Jonathon eBlake6Vladimir eBenes7Gareth eGriffiths8Olivier eNeyrolles9Geanncarlo eLugo-Villarino10Elsa eAnes11Faculdade de Farmácia, Universidade de LisboaINSERM U 1016, CNRS UMR 8104, Université Paris Descartes, Paris, France.Faculdade de Farmácia, Universidade de LisboaFaculdade de Farmácia, Universidade de LisboaInstitut de Pharmacologie et de Biologie Structurale, Université de Toulouse, Université Paul Sabatier, Toulouse, FranceFaculdade de Farmácia, Universidade de LisboaEuropean Molecular Biology LaboratoryEuropean Molecular Biology LaboratoryUniversity of Oslo, PO Box 1041, Blindern, 0316 Oslo, NorwayInstitut de Pharmacologie et de Biologie Structurale, Université de Toulouse, Université Paul Sabatier, Toulouse, FranceInstitut de Pharmacologie et de Biologie Structurale, Université de Toulouse, Université Paul Sabatier, Toulouse, FranceFaculdade de Farmácia, Universidade de LisboaMycobacterium tuberculosis (Mtb) is a successful intracellular pathogen that thrives in macrophages (Mφs). There is a need to better understand how Mtb hijacks cellular processes like phagolysosome biogenesis, a classical pathogenesis determinant. A central feature to this microbial strategy is the manipulation of actin in Mφs. Here, we examine 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 filamentation to prolong its instracellular survival inside host cells, our approach was to perform a global transcriptomic approach to assess the modulation of miRNAs upon M. smegmatis infection of the murine Mφ cell line, J774A.1. This approach yielded miR-142-3p as a top candidate to be involved in the regulation of actin dynamics required in phagocytosis. We unequivocally demonstrate 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 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 counts over time. We therefore propose the modulation of miRNAs as a novel strategy in mycobacterial infection to control factors involved in actin filamentation and other early events of phagolysosome biogenesis, such as the case presented here for miR-142-3p influencing N-Wasp activity during the phagocytosis process.http://journal.frontiersin.org/Journal/10.3389/fcimb.2013.00019/fullPhagocytosisTuberculosismiRNAmacrophagemir-142-3pN-Wasp |
| spellingShingle | Paulo eBettencourt Sabrina eMarion David ePires Leonor eSantos Claire eLastrucci Nuno eCarmo Jonathon eBlake Vladimir eBenes Gareth eGriffiths Olivier eNeyrolles Geanncarlo eLugo-Villarino Elsa eAnes 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 Frontiers in Cellular and Infection Microbiology Phagocytosis Tuberculosis miRNA macrophage mir-142-3p N-Wasp |
| 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 | Phagocytosis Tuberculosis miRNA macrophage mir-142-3p N-Wasp |
| url | http://journal.frontiersin.org/Journal/10.3389/fcimb.2013.00019/full |
| work_keys_str_mv | AT pauloebettencourt actinbindingproteinregulationbymicrornasasanovelmicrobialstrategytomodulatephagocytosisbyhostcellsthecaseofnwaspandmir1423p AT sabrinaemarion actinbindingproteinregulationbymicrornasasanovelmicrobialstrategytomodulatephagocytosisbyhostcellsthecaseofnwaspandmir1423p AT davidepires actinbindingproteinregulationbymicrornasasanovelmicrobialstrategytomodulatephagocytosisbyhostcellsthecaseofnwaspandmir1423p AT leonoresantos actinbindingproteinregulationbymicrornasasanovelmicrobialstrategytomodulatephagocytosisbyhostcellsthecaseofnwaspandmir1423p AT claireelastrucci actinbindingproteinregulationbymicrornasasanovelmicrobialstrategytomodulatephagocytosisbyhostcellsthecaseofnwaspandmir1423p AT nunoecarmo actinbindingproteinregulationbymicrornasasanovelmicrobialstrategytomodulatephagocytosisbyhostcellsthecaseofnwaspandmir1423p AT jonathoneblake actinbindingproteinregulationbymicrornasasanovelmicrobialstrategytomodulatephagocytosisbyhostcellsthecaseofnwaspandmir1423p AT vladimirebenes actinbindingproteinregulationbymicrornasasanovelmicrobialstrategytomodulatephagocytosisbyhostcellsthecaseofnwaspandmir1423p AT garethegriffiths actinbindingproteinregulationbymicrornasasanovelmicrobialstrategytomodulatephagocytosisbyhostcellsthecaseofnwaspandmir1423p AT oliviereneyrolles actinbindingproteinregulationbymicrornasasanovelmicrobialstrategytomodulatephagocytosisbyhostcellsthecaseofnwaspandmir1423p AT geanncarloelugovillarino actinbindingproteinregulationbymicrornasasanovelmicrobialstrategytomodulatephagocytosisbyhostcellsthecaseofnwaspandmir1423p AT elsaeanes actinbindingproteinregulationbymicrornasasanovelmicrobialstrategytomodulatephagocytosisbyhostcellsthecaseofnwaspandmir1423p |