Suppressor Mutations in Type II Secretion Mutants of <named-content content-type="genus-species">Vibrio cholerae</named-content>: Inactivation of the VesC Protease

ABSTRACT The type II secretion system (T2SS) is a conserved transport pathway responsible for the secretion of a range of virulence factors by many pathogens, including Vibrio cholerae. Disruption of the T2SS genes in V. cholerae results in loss of secretion, changes in cell envelope function, and g...

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Autores principales:	Chelsea S. Rule, Young-Jun Park, Jaclyn R. Delarosa, Stewart Turley, Wim G. J. Hol, Sarah McColm, Colby Gura, Frank DiMaio, Konstantin V. Korotkov, Maria Sandkvist
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	Vibrio cholerae type II secretion system serine protease suppressor protein structure Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/d20dbb7d0226484bb0f34d676bff932b
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spelling	oai:doaj.org-article:d20dbb7d0226484bb0f34d676bff932b2021-11-15T15:31:13ZSuppressor Mutations in Type II Secretion Mutants of <named-content content-type="genus-species">Vibrio cholerae</named-content>: Inactivation of the VesC Protease10.1128/mSphere.01125-202379-5042https://doaj.org/article/d20dbb7d0226484bb0f34d676bff932b2020-12-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.01125-20https://doaj.org/toc/2379-5042ABSTRACT The type II secretion system (T2SS) is a conserved transport pathway responsible for the secretion of a range of virulence factors by many pathogens, including Vibrio cholerae. Disruption of the T2SS genes in V. cholerae results in loss of secretion, changes in cell envelope function, and growth defects. While T2SS mutants are viable, high-throughput genomic analyses have listed these genes among essential genes. To investigate whether secondary mutations arise as a consequence of T2SS inactivation, we sequenced the genomes of six V. cholerae T2SS mutants with deletions or insertions in either the epsG, epsL, or epsM genes and identified secondary mutations in all mutants. Two of the six T2SS mutants contain distinct mutations in the gene encoding the T2SS-secreted protease VesC. Other mutations were found in genes coding for V. cholerae cell envelope proteins. Subsequent sequence analysis of the vesC gene in 92 additional T2SS mutant isolates identified another 19 unique mutations including insertions or deletions, sequence duplications, and single-nucleotide changes resulting in amino acid substitutions in the VesC protein. Analysis of VesC variants and the X-ray crystallographic structure of wild-type VesC suggested that all mutations lead to loss of VesC production and/or function. One possible mechanism by which V. cholerae T2SS mutagenesis can be tolerated is through selection of vesC-inactivating mutations, which may, in part, suppress cell envelope damage, establishing permissive conditions for the disruption of the T2SS. Other mutations may have been acquired in genes encoding essential cell envelope proteins to prevent proteolysis by VesC. IMPORTANCE Genome-wide transposon mutagenesis has identified the genes encoding the T2SS in Vibrio cholerae as essential for viability, but the reason for this is unclear. Mutants with deletions or insertions in these genes can be isolated, suggesting that they have acquired secondary mutations that suppress their growth defect. Through whole-genome sequencing and phenotypic analysis of T2SS mutants, we show that one means by which the growth defect can be suppressed is through mutations in the gene encoding the T2SS substrate VesC. VesC homologues are present in other Vibrio species and close relatives, and this may be why inactivation of the T2SS in species such as Vibrio vulnificus, Vibrio sp. strain 60, and Aeromonas hydrophila also results in a pleiotropic effect on their outer membrane assembly and integrity.Chelsea S. RuleYoung-Jun ParkJaclyn R. DelarosaStewart TurleyWim G. J. HolSarah McColmColby GuraFrank DiMaioKonstantin V. KorotkovMaria SandkvistAmerican Society for MicrobiologyarticleVibrio choleraetype II secretion systemserine proteasesuppressorprotein structureMicrobiologyQR1-502ENmSphere, Vol 5, Iss 6 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Vibrio cholerae type II secretion system serine protease suppressor protein structure Microbiology QR1-502
spellingShingle	Vibrio cholerae type II secretion system serine protease suppressor protein structure Microbiology QR1-502 Chelsea S. Rule Young-Jun Park Jaclyn R. Delarosa Stewart Turley Wim G. J. Hol Sarah McColm Colby Gura Frank DiMaio Konstantin V. Korotkov Maria Sandkvist Suppressor Mutations in Type II Secretion Mutants of <named-content content-type="genus-species">Vibrio cholerae</named-content>: Inactivation of the VesC Protease
description	ABSTRACT The type II secretion system (T2SS) is a conserved transport pathway responsible for the secretion of a range of virulence factors by many pathogens, including Vibrio cholerae. Disruption of the T2SS genes in V. cholerae results in loss of secretion, changes in cell envelope function, and growth defects. While T2SS mutants are viable, high-throughput genomic analyses have listed these genes among essential genes. To investigate whether secondary mutations arise as a consequence of T2SS inactivation, we sequenced the genomes of six V. cholerae T2SS mutants with deletions or insertions in either the epsG, epsL, or epsM genes and identified secondary mutations in all mutants. Two of the six T2SS mutants contain distinct mutations in the gene encoding the T2SS-secreted protease VesC. Other mutations were found in genes coding for V. cholerae cell envelope proteins. Subsequent sequence analysis of the vesC gene in 92 additional T2SS mutant isolates identified another 19 unique mutations including insertions or deletions, sequence duplications, and single-nucleotide changes resulting in amino acid substitutions in the VesC protein. Analysis of VesC variants and the X-ray crystallographic structure of wild-type VesC suggested that all mutations lead to loss of VesC production and/or function. One possible mechanism by which V. cholerae T2SS mutagenesis can be tolerated is through selection of vesC-inactivating mutations, which may, in part, suppress cell envelope damage, establishing permissive conditions for the disruption of the T2SS. Other mutations may have been acquired in genes encoding essential cell envelope proteins to prevent proteolysis by VesC. IMPORTANCE Genome-wide transposon mutagenesis has identified the genes encoding the T2SS in Vibrio cholerae as essential for viability, but the reason for this is unclear. Mutants with deletions or insertions in these genes can be isolated, suggesting that they have acquired secondary mutations that suppress their growth defect. Through whole-genome sequencing and phenotypic analysis of T2SS mutants, we show that one means by which the growth defect can be suppressed is through mutations in the gene encoding the T2SS substrate VesC. VesC homologues are present in other Vibrio species and close relatives, and this may be why inactivation of the T2SS in species such as Vibrio vulnificus, Vibrio sp. strain 60, and Aeromonas hydrophila also results in a pleiotropic effect on their outer membrane assembly and integrity.
format	article
author	Chelsea S. Rule Young-Jun Park Jaclyn R. Delarosa Stewart Turley Wim G. J. Hol Sarah McColm Colby Gura Frank DiMaio Konstantin V. Korotkov Maria Sandkvist
author_facet	Chelsea S. Rule Young-Jun Park Jaclyn R. Delarosa Stewart Turley Wim G. J. Hol Sarah McColm Colby Gura Frank DiMaio Konstantin V. Korotkov Maria Sandkvist
author_sort	Chelsea S. Rule
title	Suppressor Mutations in Type II Secretion Mutants of <named-content content-type="genus-species">Vibrio cholerae</named-content>: Inactivation of the VesC Protease
title_short	Suppressor Mutations in Type II Secretion Mutants of <named-content content-type="genus-species">Vibrio cholerae</named-content>: Inactivation of the VesC Protease
title_full	Suppressor Mutations in Type II Secretion Mutants of <named-content content-type="genus-species">Vibrio cholerae</named-content>: Inactivation of the VesC Protease
title_fullStr	Suppressor Mutations in Type II Secretion Mutants of <named-content content-type="genus-species">Vibrio cholerae</named-content>: Inactivation of the VesC Protease
title_full_unstemmed	Suppressor Mutations in Type II Secretion Mutants of <named-content content-type="genus-species">Vibrio cholerae</named-content>: Inactivation of the VesC Protease
title_sort	suppressor mutations in type ii secretion mutants of <named-content content-type="genus-species">vibrio cholerae</named-content>: inactivation of the vesc protease
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/d20dbb7d0226484bb0f34d676bff932b
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Suppressor Mutations in Type II Secretion Mutants of <named-content content-type="genus-species">Vibrio cholerae</named-content>: Inactivation of the VesC Protease

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