Meloidogyne incognita PASSE-MURAILLE (MiPM) Gene Encodes a Cell-Penetrating Protein That Interacts With the CSN5 Subunit of the COP9 Signalosome
The pathogenicity of phytonematodes relies on secreted virulence factors to rewire host cellular pathways for the benefits of the nematode. In the root-knot nematode (RKN) Meloidogyne incognita, thousands of predicted secreted proteins have been identified and are expected to interact with host prot...
Main Authors: | , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Frontiers Media S.A.
2018-06-01
|
Series: | Frontiers in Plant Science |
Subjects: | |
Online Access: | https://www.frontiersin.org/article/10.3389/fpls.2018.00904/full |
id |
doaj-8c340351f5384be1bde954c1868174f8 |
---|---|
record_format |
Article |
spelling |
doaj-8c340351f5384be1bde954c1868174f82020-11-24T21:35:05ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2018-06-01910.3389/fpls.2018.00904380609Meloidogyne incognita PASSE-MURAILLE (MiPM) Gene Encodes a Cell-Penetrating Protein That Interacts With the CSN5 Subunit of the COP9 SignalosomeCaroline Bournaud0François-Xavier Gillet1André M. Murad2Emmanuel Bresso3Erika V. S. Albuquerque4Maria F. Grossi-de-Sá5Maria F. Grossi-de-Sá6Embrapa Genetic Resources and Biotechnology, Brasília, BrazilEmbrapa Genetic Resources and Biotechnology, Brasília, BrazilEmbrapa Genetic Resources and Biotechnology, Brasília, BrazilUniversité de Lorraine, Centre National de la Recherche Scientifique, Inria, Laboratoire Lorrain de Recherche en Informatique et ses Applications, Nancy, FranceEmbrapa Genetic Resources and Biotechnology, Brasília, BrazilEmbrapa Genetic Resources and Biotechnology, Brasília, BrazilPost-Graduation Program in Genomic Science and Biotechnology, Universidade Católica de Brasília, Brasília, BrazilThe pathogenicity of phytonematodes relies on secreted virulence factors to rewire host cellular pathways for the benefits of the nematode. In the root-knot nematode (RKN) Meloidogyne incognita, thousands of predicted secreted proteins have been identified and are expected to interact with host proteins at different developmental stages of the parasite. Identifying the host targets will provide compelling evidence about the biological significance and molecular function of the predicted proteins. Here, we have focused on the hub protein CSN5, the fifth subunit of the pleiotropic and eukaryotic conserved COP9 signalosome (CSN), which is a regulatory component of the ubiquitin/proteasome system. We used affinity purification-mass spectrometry (AP-MS) to generate the interaction network of CSN5 in M. incognita-infected roots. We identified the complete CSN complex and other known CSN5 interaction partners in addition to unknown plant and M. incognita proteins. Among these, we described M. incognita PASSE-MURAILLE (MiPM), a small pioneer protein predicted to contain a secretory peptide that is up-regulated mostly in the J2 parasitic stage. We confirmed the CSN5-MiPM interaction, which occurs in the nucleus, by bimolecular fluorescence complementation (BiFC). Using MiPM as bait, a GST pull-down assay coupled with MS revealed some common protein partners between CSN5 and MiPM. We further showed by in silico and microscopic analyses that the recombinant purified MiPM protein enters the cells of Arabidopsis root tips in a non-infectious context. In further detail, the supercharged N-terminal tail of MiPM (NTT-MiPM) triggers an unknown host endocytosis pathway to penetrate the cell. The functional meaning of the CSN5-MiPM interaction in the M. incognita parasitism is discussed. Moreover, we propose that the cell-penetrating properties of some M. incognita secreted proteins might be a non-negligible mechanism for cell uptake, especially during the steps preceding the sedentary parasitic phase.https://www.frontiersin.org/article/10.3389/fpls.2018.00904/fullinteractomeroot-knot nematodeaffinity-purificationhub proteincell entryendocytosis |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Caroline Bournaud François-Xavier Gillet André M. Murad Emmanuel Bresso Erika V. S. Albuquerque Maria F. Grossi-de-Sá Maria F. Grossi-de-Sá |
spellingShingle |
Caroline Bournaud François-Xavier Gillet André M. Murad Emmanuel Bresso Erika V. S. Albuquerque Maria F. Grossi-de-Sá Maria F. Grossi-de-Sá Meloidogyne incognita PASSE-MURAILLE (MiPM) Gene Encodes a Cell-Penetrating Protein That Interacts With the CSN5 Subunit of the COP9 Signalosome Frontiers in Plant Science interactome root-knot nematode affinity-purification hub protein cell entry endocytosis |
author_facet |
Caroline Bournaud François-Xavier Gillet André M. Murad Emmanuel Bresso Erika V. S. Albuquerque Maria F. Grossi-de-Sá Maria F. Grossi-de-Sá |
author_sort |
Caroline Bournaud |
title |
Meloidogyne incognita PASSE-MURAILLE (MiPM) Gene Encodes a Cell-Penetrating Protein That Interacts With the CSN5 Subunit of the COP9 Signalosome |
title_short |
Meloidogyne incognita PASSE-MURAILLE (MiPM) Gene Encodes a Cell-Penetrating Protein That Interacts With the CSN5 Subunit of the COP9 Signalosome |
title_full |
Meloidogyne incognita PASSE-MURAILLE (MiPM) Gene Encodes a Cell-Penetrating Protein That Interacts With the CSN5 Subunit of the COP9 Signalosome |
title_fullStr |
Meloidogyne incognita PASSE-MURAILLE (MiPM) Gene Encodes a Cell-Penetrating Protein That Interacts With the CSN5 Subunit of the COP9 Signalosome |
title_full_unstemmed |
Meloidogyne incognita PASSE-MURAILLE (MiPM) Gene Encodes a Cell-Penetrating Protein That Interacts With the CSN5 Subunit of the COP9 Signalosome |
title_sort |
meloidogyne incognita passe-muraille (mipm) gene encodes a cell-penetrating protein that interacts with the csn5 subunit of the cop9 signalosome |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Plant Science |
issn |
1664-462X |
publishDate |
2018-06-01 |
description |
The pathogenicity of phytonematodes relies on secreted virulence factors to rewire host cellular pathways for the benefits of the nematode. In the root-knot nematode (RKN) Meloidogyne incognita, thousands of predicted secreted proteins have been identified and are expected to interact with host proteins at different developmental stages of the parasite. Identifying the host targets will provide compelling evidence about the biological significance and molecular function of the predicted proteins. Here, we have focused on the hub protein CSN5, the fifth subunit of the pleiotropic and eukaryotic conserved COP9 signalosome (CSN), which is a regulatory component of the ubiquitin/proteasome system. We used affinity purification-mass spectrometry (AP-MS) to generate the interaction network of CSN5 in M. incognita-infected roots. We identified the complete CSN complex and other known CSN5 interaction partners in addition to unknown plant and M. incognita proteins. Among these, we described M. incognita PASSE-MURAILLE (MiPM), a small pioneer protein predicted to contain a secretory peptide that is up-regulated mostly in the J2 parasitic stage. We confirmed the CSN5-MiPM interaction, which occurs in the nucleus, by bimolecular fluorescence complementation (BiFC). Using MiPM as bait, a GST pull-down assay coupled with MS revealed some common protein partners between CSN5 and MiPM. We further showed by in silico and microscopic analyses that the recombinant purified MiPM protein enters the cells of Arabidopsis root tips in a non-infectious context. In further detail, the supercharged N-terminal tail of MiPM (NTT-MiPM) triggers an unknown host endocytosis pathway to penetrate the cell. The functional meaning of the CSN5-MiPM interaction in the M. incognita parasitism is discussed. Moreover, we propose that the cell-penetrating properties of some M. incognita secreted proteins might be a non-negligible mechanism for cell uptake, especially during the steps preceding the sedentary parasitic phase. |
topic |
interactome root-knot nematode affinity-purification hub protein cell entry endocytosis |
url |
https://www.frontiersin.org/article/10.3389/fpls.2018.00904/full |
work_keys_str_mv |
AT carolinebournaud meloidogyneincognitapassemuraillemipmgeneencodesacellpenetratingproteinthatinteractswiththecsn5subunitofthecop9signalosome AT francoisxaviergillet meloidogyneincognitapassemuraillemipmgeneencodesacellpenetratingproteinthatinteractswiththecsn5subunitofthecop9signalosome AT andremmurad meloidogyneincognitapassemuraillemipmgeneencodesacellpenetratingproteinthatinteractswiththecsn5subunitofthecop9signalosome AT emmanuelbresso meloidogyneincognitapassemuraillemipmgeneencodesacellpenetratingproteinthatinteractswiththecsn5subunitofthecop9signalosome AT erikavsalbuquerque meloidogyneincognitapassemuraillemipmgeneencodesacellpenetratingproteinthatinteractswiththecsn5subunitofthecop9signalosome AT mariafgrossidesa meloidogyneincognitapassemuraillemipmgeneencodesacellpenetratingproteinthatinteractswiththecsn5subunitofthecop9signalosome AT mariafgrossidesa meloidogyneincognitapassemuraillemipmgeneencodesacellpenetratingproteinthatinteractswiththecsn5subunitofthecop9signalosome |
_version_ |
1725946719374409728 |