Genetic Reporter System for Positioning of Proteins at the Bacterial Pole

ABSTRACT Spatial organization within bacteria is fundamental to many cellular processes, although the basic mechanisms underlying localization of proteins to specific sites within bacteria are poorly understood. The study of protein positioning has been limited by a paucity of methods that allow rap...

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Autores principales:	Kathryn R. Fixen, Anuradha Janakiraman, Sean Garrity, Daniel J. Slade, Andrew N. Gray, Nilay Karahan, Ann Hochschild, Marcia B. Goldberg
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2012
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/02d8233260684148abdd8a078616344b
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spelling	oai:doaj.org-article:02d8233260684148abdd8a078616344b2021-11-15T15:39:09ZGenetic Reporter System for Positioning of Proteins at the Bacterial Pole10.1128/mBio.00251-112150-7511https://doaj.org/article/02d8233260684148abdd8a078616344b2012-05-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00251-11https://doaj.org/toc/2150-7511ABSTRACT Spatial organization within bacteria is fundamental to many cellular processes, although the basic mechanisms underlying localization of proteins to specific sites within bacteria are poorly understood. The study of protein positioning has been limited by a paucity of methods that allow rapid large-scale screening for mutants in which protein positioning is altered. We developed a genetic reporter system for protein localization to the pole within the bacterial cytoplasm that allows saturation screening for mutants in Escherichia coli in which protein localization is altered. Utilizing this system, we identify proteins required for proper positioning of the Shigella autotransporter IcsA. Autotransporters, widely distributed bacterial virulence proteins, are secreted at the bacterial pole. We show that the conserved cell division protein FtsQ is required for localization of IcsA and other autotransporters to the pole. We demonstrate further that this system can be applied to the study of proteins other than autotransporters that display polar positioning within bacterial cells. IMPORTANCE Many proteins localize to specific sites within bacterial cells, and localization to these sites is frequently critical to proper protein function. The mechanisms that underlie protein localization are incompletely understood, in part because of the paucity of methods that allow saturation screening for mutants in which protein localization is altered. We developed a genetic reporter assay that enables screening of bacterial populations for changes in localization of proteins to the bacterial pole, and we demonstrate the utility of the system in identifying factors required for proper localization of the polar Shigella autotransporter protein IcsA. Using this method, we identify the conserved cell division protein FtsQ as being required for positioning of IcsA to the bacterial pole. We demonstrate further that the requirement for FtsQ for polar positioning applies to other autotransporters and that the method can be applied to polar proteins other than autotransporters.Kathryn R. FixenAnuradha JanakiramanSean GarrityDaniel J. SladeAndrew N. GrayNilay KarahanAnn HochschildMarcia B. GoldbergAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 3, Iss 2 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Kathryn R. Fixen Anuradha Janakiraman Sean Garrity Daniel J. Slade Andrew N. Gray Nilay Karahan Ann Hochschild Marcia B. Goldberg Genetic Reporter System for Positioning of Proteins at the Bacterial Pole
description	ABSTRACT Spatial organization within bacteria is fundamental to many cellular processes, although the basic mechanisms underlying localization of proteins to specific sites within bacteria are poorly understood. The study of protein positioning has been limited by a paucity of methods that allow rapid large-scale screening for mutants in which protein positioning is altered. We developed a genetic reporter system for protein localization to the pole within the bacterial cytoplasm that allows saturation screening for mutants in Escherichia coli in which protein localization is altered. Utilizing this system, we identify proteins required for proper positioning of the Shigella autotransporter IcsA. Autotransporters, widely distributed bacterial virulence proteins, are secreted at the bacterial pole. We show that the conserved cell division protein FtsQ is required for localization of IcsA and other autotransporters to the pole. We demonstrate further that this system can be applied to the study of proteins other than autotransporters that display polar positioning within bacterial cells. IMPORTANCE Many proteins localize to specific sites within bacterial cells, and localization to these sites is frequently critical to proper protein function. The mechanisms that underlie protein localization are incompletely understood, in part because of the paucity of methods that allow saturation screening for mutants in which protein localization is altered. We developed a genetic reporter assay that enables screening of bacterial populations for changes in localization of proteins to the bacterial pole, and we demonstrate the utility of the system in identifying factors required for proper localization of the polar Shigella autotransporter protein IcsA. Using this method, we identify the conserved cell division protein FtsQ as being required for positioning of IcsA to the bacterial pole. We demonstrate further that the requirement for FtsQ for polar positioning applies to other autotransporters and that the method can be applied to polar proteins other than autotransporters.
format	article
author	Kathryn R. Fixen Anuradha Janakiraman Sean Garrity Daniel J. Slade Andrew N. Gray Nilay Karahan Ann Hochschild Marcia B. Goldberg
author_facet	Kathryn R. Fixen Anuradha Janakiraman Sean Garrity Daniel J. Slade Andrew N. Gray Nilay Karahan Ann Hochschild Marcia B. Goldberg
author_sort	Kathryn R. Fixen
title	Genetic Reporter System for Positioning of Proteins at the Bacterial Pole
title_short	Genetic Reporter System for Positioning of Proteins at the Bacterial Pole
title_full	Genetic Reporter System for Positioning of Proteins at the Bacterial Pole
title_fullStr	Genetic Reporter System for Positioning of Proteins at the Bacterial Pole
title_full_unstemmed	Genetic Reporter System for Positioning of Proteins at the Bacterial Pole
title_sort	genetic reporter system for positioning of proteins at the bacterial pole
publisher	American Society for Microbiology
publishDate	2012
url	https://doaj.org/article/02d8233260684148abdd8a078616344b
work_keys_str_mv	AT kathrynrfixen geneticreportersystemforpositioningofproteinsatthebacterialpole AT anuradhajanakiraman geneticreportersystemforpositioningofproteinsatthebacterialpole AT seangarrity geneticreportersystemforpositioningofproteinsatthebacterialpole AT danieljslade geneticreportersystemforpositioningofproteinsatthebacterialpole AT andrewngray geneticreportersystemforpositioningofproteinsatthebacterialpole AT nilaykarahan geneticreportersystemforpositioningofproteinsatthebacterialpole AT annhochschild geneticreportersystemforpositioningofproteinsatthebacterialpole AT marciabgoldberg geneticreportersystemforpositioningofproteinsatthebacterialpole
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Genetic Reporter System for Positioning of Proteins at the Bacterial Pole

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