Glutathionylation of <italic toggle="yes">Yersinia pestis</italic> LcrV and Its Effects on Plague Pathogenesis

ABSTRACT Glutathionylation, the formation of reversible mixed disulfides between glutathione and protein cysteine residues, is a posttranslational modification previously observed for intracellular proteins of bacteria. Here we show that Yersinia pestis LcrV, a secreted protein capping the type III...

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Autores principales:	Anthony Mitchell, Christina Tam, Derek Elli, Thomas Charlton, Patrick Osei-Owusu, Farbod Fazlollahi, Kym F. Faull, Olaf Schneewind
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2017
Materias:	macrophage Yersinia pestis glutathionylation innate immunity plague ribosomal protein S3 (RPS3) Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/454ceb1dc8484d7fba68baeebe5881d0
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spelling	oai:doaj.org-article:454ceb1dc8484d7fba68baeebe5881d02021-11-15T15:51:30ZGlutathionylation of <italic toggle="yes">Yersinia pestis</italic> LcrV and Its Effects on Plague Pathogenesis10.1128/mBio.00646-172150-7511https://doaj.org/article/454ceb1dc8484d7fba68baeebe5881d02017-07-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00646-17https://doaj.org/toc/2150-7511ABSTRACT Glutathionylation, the formation of reversible mixed disulfides between glutathione and protein cysteine residues, is a posttranslational modification previously observed for intracellular proteins of bacteria. Here we show that Yersinia pestis LcrV, a secreted protein capping the type III secretion machine, is glutathionylated at Cys273 and that this modification promotes association with host ribosomal protein S3 (RPS3), moderates Y. pestis type III effector transport and killing of macrophages, and enhances bubonic plague pathogenesis in mice and rats. Secreted LcrV was purified and analyzed by mass spectrometry to reveal glutathionylation, a modification that is abolished by the codon substitution Cys273Ala in lcrV. Moreover, the lcrVC273A mutation enhanced the survival of animals in models of bubonic plague. Investigating the molecular mechanism responsible for these virulence attributes, we identified macrophage RPS3 as a ligand of LcrV, an association that is perturbed by the Cys273Ala substitution. Furthermore, macrophages infected by the lcrVC273A variant displayed accelerated apoptotic death and diminished proinflammatory cytokine release. Deletion of gshB, which encodes glutathione synthetase of Y. pestis, resulted in undetectable levels of intracellular glutathione, and we used a Y. pestis ΔgshB mutant to characterize the biochemical pathway of LcrV glutathionylation, establishing that LcrV is modified after its transport to the type III needle via disulfide bond formation with extracellular oxidized glutathione. IMPORTANCE Yersinia pestis, the causative agent of plague, has killed large segments of the human population; however, the molecular bases for the extraordinary virulence attributes of this pathogen are not well understood. We show here that LcrV, the cap protein of bacterial type III secretion needles, is modified by host glutathione and that this modification contributes to the high virulence of Y. pestis in mouse and rat models for bubonic plague. These data suggest that Y. pestis exploits glutathione in host tissues to activate a virulence strategy, thereby accelerating plague pathogenesis.Anthony MitchellChristina TamDerek ElliThomas CharltonPatrick Osei-OwusuFarbod FazlollahiKym F. FaullOlaf SchneewindAmerican Society for MicrobiologyarticlemacrophageYersinia pestisglutathionylationinnate immunityplagueribosomal protein S3 (RPS3)MicrobiologyQR1-502ENmBio, Vol 8, Iss 3 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	macrophage Yersinia pestis glutathionylation innate immunity plague ribosomal protein S3 (RPS3) Microbiology QR1-502
spellingShingle	macrophage Yersinia pestis glutathionylation innate immunity plague ribosomal protein S3 (RPS3) Microbiology QR1-502 Anthony Mitchell Christina Tam Derek Elli Thomas Charlton Patrick Osei-Owusu Farbod Fazlollahi Kym F. Faull Olaf Schneewind Glutathionylation of <italic toggle="yes">Yersinia pestis</italic> LcrV and Its Effects on Plague Pathogenesis
description	ABSTRACT Glutathionylation, the formation of reversible mixed disulfides between glutathione and protein cysteine residues, is a posttranslational modification previously observed for intracellular proteins of bacteria. Here we show that Yersinia pestis LcrV, a secreted protein capping the type III secretion machine, is glutathionylated at Cys273 and that this modification promotes association with host ribosomal protein S3 (RPS3), moderates Y. pestis type III effector transport and killing of macrophages, and enhances bubonic plague pathogenesis in mice and rats. Secreted LcrV was purified and analyzed by mass spectrometry to reveal glutathionylation, a modification that is abolished by the codon substitution Cys273Ala in lcrV. Moreover, the lcrVC273A mutation enhanced the survival of animals in models of bubonic plague. Investigating the molecular mechanism responsible for these virulence attributes, we identified macrophage RPS3 as a ligand of LcrV, an association that is perturbed by the Cys273Ala substitution. Furthermore, macrophages infected by the lcrVC273A variant displayed accelerated apoptotic death and diminished proinflammatory cytokine release. Deletion of gshB, which encodes glutathione synthetase of Y. pestis, resulted in undetectable levels of intracellular glutathione, and we used a Y. pestis ΔgshB mutant to characterize the biochemical pathway of LcrV glutathionylation, establishing that LcrV is modified after its transport to the type III needle via disulfide bond formation with extracellular oxidized glutathione. IMPORTANCE Yersinia pestis, the causative agent of plague, has killed large segments of the human population; however, the molecular bases for the extraordinary virulence attributes of this pathogen are not well understood. We show here that LcrV, the cap protein of bacterial type III secretion needles, is modified by host glutathione and that this modification contributes to the high virulence of Y. pestis in mouse and rat models for bubonic plague. These data suggest that Y. pestis exploits glutathione in host tissues to activate a virulence strategy, thereby accelerating plague pathogenesis.
format	article
author	Anthony Mitchell Christina Tam Derek Elli Thomas Charlton Patrick Osei-Owusu Farbod Fazlollahi Kym F. Faull Olaf Schneewind
author_facet	Anthony Mitchell Christina Tam Derek Elli Thomas Charlton Patrick Osei-Owusu Farbod Fazlollahi Kym F. Faull Olaf Schneewind
author_sort	Anthony Mitchell
title	Glutathionylation of <italic toggle="yes">Yersinia pestis</italic> LcrV and Its Effects on Plague Pathogenesis
title_short	Glutathionylation of <italic toggle="yes">Yersinia pestis</italic> LcrV and Its Effects on Plague Pathogenesis
title_full	Glutathionylation of <italic toggle="yes">Yersinia pestis</italic> LcrV and Its Effects on Plague Pathogenesis
title_fullStr	Glutathionylation of <italic toggle="yes">Yersinia pestis</italic> LcrV and Its Effects on Plague Pathogenesis
title_full_unstemmed	Glutathionylation of <italic toggle="yes">Yersinia pestis</italic> LcrV and Its Effects on Plague Pathogenesis
title_sort	glutathionylation of <italic toggle="yes">yersinia pestis</italic> lcrv and its effects on plague pathogenesis
publisher	American Society for Microbiology
publishDate	2017
url	https://doaj.org/article/454ceb1dc8484d7fba68baeebe5881d0
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Glutathionylation of <italic toggle="yes">Yersinia pestis</italic> LcrV and Its Effects on Plague Pathogenesis

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