A Two-Component Regulatory System Impacts Extracellular Membrane-Derived Vesicle Production in Group A Streptococcus
ABSTRACT Export of macromolecules via extracellular membrane-derived vesicles (MVs) plays an important role in the biology of Gram-negative bacteria. Gram-positive bacteria have also recently been reported to produce MVs; however, the composition and mechanisms governing vesiculogenesis in Gram-posi...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
American Society for Microbiology
2016-12-01
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| Series: | mBio |
| Online Access: | https://journals.asm.org/doi/10.1128/mBio.00207-16 |
| _version_ | 1818434462551113728 |
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| author | Ulrike Resch James Anthony Tsatsaronis Anaïs Le Rhun Gerald Stübiger Manfred Rohde Sergo Kasvandik Susanne Holzmeister Philip Tinnefeld Sun Nyunt Wai Emmanuelle Charpentier |
| author_facet | Ulrike Resch James Anthony Tsatsaronis Anaïs Le Rhun Gerald Stübiger Manfred Rohde Sergo Kasvandik Susanne Holzmeister Philip Tinnefeld Sun Nyunt Wai Emmanuelle Charpentier |
| author_sort | Ulrike Resch |
| collection | DOAJ |
| description | ABSTRACT Export of macromolecules via extracellular membrane-derived vesicles (MVs) plays an important role in the biology of Gram-negative bacteria. Gram-positive bacteria have also recently been reported to produce MVs; however, the composition and mechanisms governing vesiculogenesis in Gram-positive bacteria remain undefined. Here, we describe MV production in the Gram-positive human pathogen group A streptococcus (GAS), the etiological agent of necrotizing fasciitis and streptococcal toxic shock syndrome. M1 serotype GAS isolates in culture exhibit MV structures both on the cell wall surface and in the near vicinity of bacterial cells. A comprehensive analysis of MV proteins identified both virulence-associated protein substrates of the general secretory pathway in addition to “anchorless surface proteins.” Characteristic differences in the contents, distributions, and fatty acid compositions of specific lipids between MVs and GAS cell membrane were also observed. Furthermore, deep RNA sequencing of vesicular RNAs revealed that GAS MVs contained differentially abundant RNA species relative to bacterial cellular RNA. MV production by GAS strains varied in a manner dependent on an intact two-component system, CovRS, with MV production negatively regulated by the system. Modulation of MV production through CovRS was found to be independent of both GAS cysteine protease SpeB and capsule biosynthesis. Our data provide an explanation for GAS secretion of macromolecules, including RNAs, lipids, and proteins, and illustrate a regulatory mechanism coordinating this secretory response. IMPORTANCE Group A streptococcus (GAS) is a Gram-positive bacterial pathogen responsible for more than 500,000 deaths annually. Establishment of GAS infection is dependent on a suite of proteins exported via the general secretory pathway. Here, we show that GAS naturally produces extracellular vesicles with a unique lipid composition that are laden with proteins and RNAs. Interestingly, both virulence-associated proteins and RNA species were found to be differentially abundant in vesicles relative to the bacteria. Furthermore, we show that genetic disruption of the virulence-associated two-component regulator CovRS leads to an increase in vesicle production. This study comprehensively describes the protein, RNA, and lipid composition of GAS-secreted MVs and alludes to a regulatory system impacting this process. |
| first_indexed | 2024-12-14T16:37:22Z |
| format | Article |
| id | doaj.art-0ec16a69cbcb49f891c986aa591fca62 |
| institution | Directory Open Access Journal |
| issn | 2150-7511 |
| language | English |
| last_indexed | 2024-12-14T16:37:22Z |
| publishDate | 2016-12-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mBio |
| spelling | doaj.art-0ec16a69cbcb49f891c986aa591fca622022-12-21T22:54:24ZengAmerican Society for MicrobiologymBio2150-75112016-12-017610.1128/mBio.00207-16A Two-Component Regulatory System Impacts Extracellular Membrane-Derived Vesicle Production in Group A StreptococcusUlrike Resch0James Anthony Tsatsaronis1Anaïs Le Rhun2Gerald Stübiger3Manfred Rohde4Sergo Kasvandik5Susanne Holzmeister6Philip Tinnefeld7Sun Nyunt Wai8Emmanuelle Charpentier9The Laboratory for Molecular Infection Medicine Sweden (MIMS), Department of Molecular Biology, Umeå University, Umeå, SwedenDepartment of Regulation in Infection Biology, Helmholtz Centre for Infection Research, Braunschweig, GermanyThe Laboratory for Molecular Infection Medicine Sweden (MIMS), Department of Molecular Biology, Umeå University, Umeå, SwedenDepartment of Biomedical Imaging and Image-Guided Therapy, Medical University, Vienna, Vienna, AustriaCentral Facility for Microscopy, Helmholtz Centre for Infection Research, Braunschweig, GermanyUniversity of Tartu, Institute of Technology, Tartu, EstoniaNanoBioScience Group, Institute for Physical and Theoretical Chemistry, TU, Braunschweig, Braunschweig, GermanyNanoBioScience Group, Institute for Physical and Theoretical Chemistry, TU, Braunschweig, Braunschweig, GermanyThe Laboratory for Molecular Infection Medicine Sweden (MIMS), Department of Molecular Biology, Umeå University, Umeå, SwedenThe Laboratory for Molecular Infection Medicine Sweden (MIMS), Department of Molecular Biology, Umeå University, Umeå, SwedenABSTRACT Export of macromolecules via extracellular membrane-derived vesicles (MVs) plays an important role in the biology of Gram-negative bacteria. Gram-positive bacteria have also recently been reported to produce MVs; however, the composition and mechanisms governing vesiculogenesis in Gram-positive bacteria remain undefined. Here, we describe MV production in the Gram-positive human pathogen group A streptococcus (GAS), the etiological agent of necrotizing fasciitis and streptococcal toxic shock syndrome. M1 serotype GAS isolates in culture exhibit MV structures both on the cell wall surface and in the near vicinity of bacterial cells. A comprehensive analysis of MV proteins identified both virulence-associated protein substrates of the general secretory pathway in addition to “anchorless surface proteins.” Characteristic differences in the contents, distributions, and fatty acid compositions of specific lipids between MVs and GAS cell membrane were also observed. Furthermore, deep RNA sequencing of vesicular RNAs revealed that GAS MVs contained differentially abundant RNA species relative to bacterial cellular RNA. MV production by GAS strains varied in a manner dependent on an intact two-component system, CovRS, with MV production negatively regulated by the system. Modulation of MV production through CovRS was found to be independent of both GAS cysteine protease SpeB and capsule biosynthesis. Our data provide an explanation for GAS secretion of macromolecules, including RNAs, lipids, and proteins, and illustrate a regulatory mechanism coordinating this secretory response. IMPORTANCE Group A streptococcus (GAS) is a Gram-positive bacterial pathogen responsible for more than 500,000 deaths annually. Establishment of GAS infection is dependent on a suite of proteins exported via the general secretory pathway. Here, we show that GAS naturally produces extracellular vesicles with a unique lipid composition that are laden with proteins and RNAs. Interestingly, both virulence-associated proteins and RNA species were found to be differentially abundant in vesicles relative to the bacteria. Furthermore, we show that genetic disruption of the virulence-associated two-component regulator CovRS leads to an increase in vesicle production. This study comprehensively describes the protein, RNA, and lipid composition of GAS-secreted MVs and alludes to a regulatory system impacting this process.https://journals.asm.org/doi/10.1128/mBio.00207-16 |
| spellingShingle | Ulrike Resch James Anthony Tsatsaronis Anaïs Le Rhun Gerald Stübiger Manfred Rohde Sergo Kasvandik Susanne Holzmeister Philip Tinnefeld Sun Nyunt Wai Emmanuelle Charpentier A Two-Component Regulatory System Impacts Extracellular Membrane-Derived Vesicle Production in Group A Streptococcus mBio |
| title | A Two-Component Regulatory System Impacts Extracellular Membrane-Derived Vesicle Production in Group A Streptococcus |
| title_full | A Two-Component Regulatory System Impacts Extracellular Membrane-Derived Vesicle Production in Group A Streptococcus |
| title_fullStr | A Two-Component Regulatory System Impacts Extracellular Membrane-Derived Vesicle Production in Group A Streptococcus |
| title_full_unstemmed | A Two-Component Regulatory System Impacts Extracellular Membrane-Derived Vesicle Production in Group A Streptococcus |
| title_short | A Two-Component Regulatory System Impacts Extracellular Membrane-Derived Vesicle Production in Group A Streptococcus |
| title_sort | two component regulatory system impacts extracellular membrane derived vesicle production in group a streptococcus |
| url | https://journals.asm.org/doi/10.1128/mBio.00207-16 |
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