Essential Role of the ESX-5 Secretion System in Outer Membrane Permeability of Pathogenic Mycobacteria

Handle URI:
http://hdl.handle.net/10754/552778
Title:
Essential Role of the ESX-5 Secretion System in Outer Membrane Permeability of Pathogenic Mycobacteria
Authors:
Ates, Louis S.; Ummels, Roy; Commandeur, Susanna; van der Weerd, Robert; Sparrius, Marion; Weerdenburg, Eveline; Alber, Marina; Kalscheuer, Rainer; Piersma, Sander R.; Abdallah, Abdallah; Abd El Ghany, Moataz; Abdel-Haleem, Alyaa M.; Pain, Arnab ( 0000-0002-1755-2819 ) ; Jiménez, Connie R.; Bitter, Wilbert; Houben, Edith N.G.
Abstract:
Mycobacteria possess different type VII secretion (T7S) systems to secrete proteins across their unusual cell envelope. One of these systems, ESX-5, is only present in slow-growing mycobacteria and responsible for the secretion of multiple substrates. However, the role of ESX-5 substrates in growth and/or virulence is largely unknown. In this study, we show that esx-5 is essential for growth of both Mycobacterium marinum and Mycobacterium bovis. Remarkably, this essentiality can be rescued by increasing the permeability of the outer membrane, either by altering its lipid composition or by the introduction of the heterologous porin MspA. Mutagenesis of the first nucleotide-binding domain of the membrane ATPase EccC5 prevented both ESX-5-dependent secretion and bacterial growth, but did not affect ESX-5 complex assembly. This suggests that the rescuing effect is not due to pores formed by the ESX-5 membrane complex, but caused by ESX-5 activity. Subsequent proteomic analysis to identify crucial ESX-5 substrates confirmed that all detectable PE and PPE proteins in the cell surface and cell envelope fractions were routed through ESX-5. Additionally, saturated transposon-directed insertion-site sequencing (TraDIS) was applied to both wild-type M. marinum cells and cells expressing mspA to identify genes that are not essential anymore in the presence of MspA. This analysis confirmed the importance of esx-5, but we could not identify essential ESX-5 substrates, indicating that multiple of these substrates are together responsible for the essentiality. Finally, examination of phenotypes on defined carbon sources revealed that an esx-5 mutant is strongly impaired in the uptake and utilization of hydrophobic carbon sources. Based on these data, we propose a model in which the ESX-5 system is responsible for the transport of cell envelope proteins that are required for nutrient uptake. These proteins might in this way compensate for the lack of MspA-like porins in slow-growing mycobacteria.
KAUST Department:
Biological and Environmental Sciences and Engineering (BESE) Division
Citation:
Essential Role of the ESX-5 Secretion System in Outer Membrane Permeability of Pathogenic Mycobacteria 2015, 11 (5):e1005190 PLOS Genetics
Publisher:
Public Library of Science (PLoS)
Journal:
PLOS Genetics
Issue Date:
4-May-2015
DOI:
10.1371/journal.pgen.1005190
PubMed ID:
25938982
PubMed Central ID:
PMC4418733
Type:
Article
ISSN:
1553-7404
Additional Links:
http://dx.plos.org/10.1371/journal.pgen.1005190
Appears in Collections:
Articles; Biological and Environmental Sciences and Engineering (BESE) Division

Full metadata record

DC FieldValue Language
dc.contributor.authorAtes, Louis S.en
dc.contributor.authorUmmels, Royen
dc.contributor.authorCommandeur, Susannaen
dc.contributor.authorvan der Weerd, Roberten
dc.contributor.authorSparrius, Marionen
dc.contributor.authorWeerdenburg, Evelineen
dc.contributor.authorAlber, Marinaen
dc.contributor.authorKalscheuer, Raineren
dc.contributor.authorPiersma, Sander R.en
dc.contributor.authorAbdallah, Abdallahen
dc.contributor.authorAbd El Ghany, Moatazen
dc.contributor.authorAbdel-Haleem, Alyaa M.en
dc.contributor.authorPain, Arnaben
dc.contributor.authorJiménez, Connie R.en
dc.contributor.authorBitter, Wilberten
dc.contributor.authorHouben, Edith N.G.en
dc.date.accessioned2015-05-14T07:05:47Zen
dc.date.available2015-05-14T07:05:47Zen
dc.date.issued2015-05-04en
dc.identifier.citationEssential Role of the ESX-5 Secretion System in Outer Membrane Permeability of Pathogenic Mycobacteria 2015, 11 (5):e1005190 PLOS Geneticsen
dc.identifier.issn1553-7404en
dc.identifier.pmid25938982en
dc.identifier.doi10.1371/journal.pgen.1005190en
dc.identifier.urihttp://hdl.handle.net/10754/552778en
dc.description.abstractMycobacteria possess different type VII secretion (T7S) systems to secrete proteins across their unusual cell envelope. One of these systems, ESX-5, is only present in slow-growing mycobacteria and responsible for the secretion of multiple substrates. However, the role of ESX-5 substrates in growth and/or virulence is largely unknown. In this study, we show that esx-5 is essential for growth of both Mycobacterium marinum and Mycobacterium bovis. Remarkably, this essentiality can be rescued by increasing the permeability of the outer membrane, either by altering its lipid composition or by the introduction of the heterologous porin MspA. Mutagenesis of the first nucleotide-binding domain of the membrane ATPase EccC5 prevented both ESX-5-dependent secretion and bacterial growth, but did not affect ESX-5 complex assembly. This suggests that the rescuing effect is not due to pores formed by the ESX-5 membrane complex, but caused by ESX-5 activity. Subsequent proteomic analysis to identify crucial ESX-5 substrates confirmed that all detectable PE and PPE proteins in the cell surface and cell envelope fractions were routed through ESX-5. Additionally, saturated transposon-directed insertion-site sequencing (TraDIS) was applied to both wild-type M. marinum cells and cells expressing mspA to identify genes that are not essential anymore in the presence of MspA. This analysis confirmed the importance of esx-5, but we could not identify essential ESX-5 substrates, indicating that multiple of these substrates are together responsible for the essentiality. Finally, examination of phenotypes on defined carbon sources revealed that an esx-5 mutant is strongly impaired in the uptake and utilization of hydrophobic carbon sources. Based on these data, we propose a model in which the ESX-5 system is responsible for the transport of cell envelope proteins that are required for nutrient uptake. These proteins might in this way compensate for the lack of MspA-like porins in slow-growing mycobacteria.en
dc.publisherPublic Library of Science (PLoS)en
dc.relation.urlhttp://dx.plos.org/10.1371/journal.pgen.1005190en
dc.rightsThis is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. http://creativecommons.org/licenses/by/4.0/en
dc.titleEssential Role of the ESX-5 Secretion System in Outer Membrane Permeability of Pathogenic Mycobacteriaen
dc.typeArticleen
dc.contributor.departmentBiological and Environmental Sciences and Engineering (BESE) Divisionen
dc.identifier.journalPLOS Geneticsen
dc.identifier.pmcidPMC4418733en
dc.eprint.versionPublisher's Version/PDFen
dc.contributor.institutionDepartment of Medical Microbiology and Infection Control, VU University Medical Center, Amsterdam, the Netherlandsen
dc.contributor.institutionInstitute for Medical Microbiology and Hospital Hygiene, Heinrich-Heine-University Düsseldorf, Düsseldorf, Germanyen
dc.contributor.institutionDepartment of Medical Oncology, OncoProteomics Laboratory, VU University Medical Center, Amsterdam, the Netherlandsen
dc.contributor.institutionSection Molecular Microbiology, Amsterdam Institute of Molecules, Medicine & Systems, Vrije Universiteit Amsterdam, Amsterdam, the Netherlandsen
kaust.authorAbd El Ghany, Moatazen
kaust.authorPain, Arnaben
kaust.authorAbdallah, Abdallahen
kaust.authorAbdel-Haleem, Alyaa M.en

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