The EXIT Strategy: an Approach for Identifying Bacterial Proteins Exported during Host Infection

ABSTRACT Exported proteins of bacterial pathogens function both in essential physiological processes and in virulence. Past efforts to identify exported proteins were limited by the use of bacteria growing under laboratory (in vitro) conditions. Thus, exported proteins that are exported only or pref...

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Autores principales:	E. F. Perkowski, K. E. Zulauf, D. Weerakoon, J. D. Hayden, T. R. Ioerger, D. Oreper, S. M. Gomez, J. C. Sacchettini, M. Braunstein
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2017
Materias:	beta-lactamase reporter EXIT Mycobacterium tuberculosis in vivo membrane proteins protein export Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/334d92456bd24feca57bbc688b7895ba
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spelling	oai:doaj.org-article:334d92456bd24feca57bbc688b7895ba2021-11-15T15:51:00ZThe EXIT Strategy: an Approach for Identifying Bacterial Proteins Exported during Host Infection10.1128/mBio.00333-172150-7511https://doaj.org/article/334d92456bd24feca57bbc688b7895ba2017-05-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00333-17https://doaj.org/toc/2150-7511ABSTRACT Exported proteins of bacterial pathogens function both in essential physiological processes and in virulence. Past efforts to identify exported proteins were limited by the use of bacteria growing under laboratory (in vitro) conditions. Thus, exported proteins that are exported only or preferentially in the context of infection may be overlooked. To solve this problem, we developed a genome-wide method, named EXIT (exported in vivotechnology), to identify proteins that are exported by bacteria during infection and applied it to Mycobacterium tuberculosis during murine infection. Our studies validate the power of EXIT to identify proteins exported during infection on an unprecedented scale (593 proteins) and to reveal in vivo induced exported proteins (i.e., proteins exported significantly more during in vivo infection than in vitro). Our EXIT data also provide an unmatched resource for mapping the topology of M. tuberculosis membrane proteins. As a new approach for identifying exported proteins, EXIT has potential applicability to other pathogens and experimental conditions. IMPORTANCE There is long-standing interest in identifying exported proteins of bacteria as they play critical roles in physiology and virulence and are commonly immunogenic antigens and targets of antibiotics. While significant effort has been made to identify the bacterial proteins that are exported beyond the cytoplasm to the membrane, cell wall, or host environment, current methods to identify exported proteins are limited by their use of bacteria growing under laboratory (in vitro) conditions. Because in vitro conditions do not mimic the complexity of the host environment, critical exported proteins that are preferentially exported in the context of infection may be overlooked. We developed a novel method to identify proteins that are exported by bacteria during host infection and applied it to identify Mycobacterium tuberculosis proteins exported in a mouse model of tuberculosis.E. F. PerkowskiK. E. ZulaufD. WeerakoonJ. D. HaydenT. R. IoergerD. OreperS. M. GomezJ. C. SacchettiniM. BraunsteinAmerican Society for Microbiologyarticlebeta-lactamase reporterEXITMycobacterium tuberculosisin vivomembrane proteinsprotein exportMicrobiologyQR1-502ENmBio, Vol 8, Iss 2 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	beta-lactamase reporter EXIT Mycobacterium tuberculosis in vivo membrane proteins protein export Microbiology QR1-502
spellingShingle	beta-lactamase reporter EXIT Mycobacterium tuberculosis in vivo membrane proteins protein export Microbiology QR1-502 E. F. Perkowski K. E. Zulauf D. Weerakoon J. D. Hayden T. R. Ioerger D. Oreper S. M. Gomez J. C. Sacchettini M. Braunstein The EXIT Strategy: an Approach for Identifying Bacterial Proteins Exported during Host Infection
description	ABSTRACT Exported proteins of bacterial pathogens function both in essential physiological processes and in virulence. Past efforts to identify exported proteins were limited by the use of bacteria growing under laboratory (in vitro) conditions. Thus, exported proteins that are exported only or preferentially in the context of infection may be overlooked. To solve this problem, we developed a genome-wide method, named EXIT (exported in vivotechnology), to identify proteins that are exported by bacteria during infection and applied it to Mycobacterium tuberculosis during murine infection. Our studies validate the power of EXIT to identify proteins exported during infection on an unprecedented scale (593 proteins) and to reveal in vivo induced exported proteins (i.e., proteins exported significantly more during in vivo infection than in vitro). Our EXIT data also provide an unmatched resource for mapping the topology of M. tuberculosis membrane proteins. As a new approach for identifying exported proteins, EXIT has potential applicability to other pathogens and experimental conditions. IMPORTANCE There is long-standing interest in identifying exported proteins of bacteria as they play critical roles in physiology and virulence and are commonly immunogenic antigens and targets of antibiotics. While significant effort has been made to identify the bacterial proteins that are exported beyond the cytoplasm to the membrane, cell wall, or host environment, current methods to identify exported proteins are limited by their use of bacteria growing under laboratory (in vitro) conditions. Because in vitro conditions do not mimic the complexity of the host environment, critical exported proteins that are preferentially exported in the context of infection may be overlooked. We developed a novel method to identify proteins that are exported by bacteria during host infection and applied it to identify Mycobacterium tuberculosis proteins exported in a mouse model of tuberculosis.
format	article
author	E. F. Perkowski K. E. Zulauf D. Weerakoon J. D. Hayden T. R. Ioerger D. Oreper S. M. Gomez J. C. Sacchettini M. Braunstein
author_facet	E. F. Perkowski K. E. Zulauf D. Weerakoon J. D. Hayden T. R. Ioerger D. Oreper S. M. Gomez J. C. Sacchettini M. Braunstein
author_sort	E. F. Perkowski
title	The EXIT Strategy: an Approach for Identifying Bacterial Proteins Exported during Host Infection
title_short	The EXIT Strategy: an Approach for Identifying Bacterial Proteins Exported during Host Infection
title_full	The EXIT Strategy: an Approach for Identifying Bacterial Proteins Exported during Host Infection
title_fullStr	The EXIT Strategy: an Approach for Identifying Bacterial Proteins Exported during Host Infection
title_full_unstemmed	The EXIT Strategy: an Approach for Identifying Bacterial Proteins Exported during Host Infection
title_sort	exit strategy: an approach for identifying bacterial proteins exported during host infection
publisher	American Society for Microbiology
publishDate	2017
url	https://doaj.org/article/334d92456bd24feca57bbc688b7895ba
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The EXIT Strategy: an Approach for Identifying Bacterial Proteins Exported during Host Infection

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