Bacteria Getting into Shape: Genetic Determinants of <italic toggle="yes">E. coli</italic> Morphology

ABSTRACT Perturbation of cellular processes is a prevailing approach to understanding biology. To better understand the complicated biology that defines bacterial shape, a sensitive, high-content platform was developed to detect multiple morphological defect phenotypes using microscopy. We examined...

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Autores principales:	Shawn French, Jean-Philippe Côté, Jonathan M. Stokes, Ray Truant, Eric D. Brown
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2017
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/91f5ed2ea2764533861ff05194d6953f
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spelling	oai:doaj.org-article:91f5ed2ea2764533861ff05194d6953f2021-11-15T15:51:00ZBacteria Getting into Shape: Genetic Determinants of <italic toggle="yes">E. coli</italic> Morphology10.1128/mBio.01977-162150-7511https://doaj.org/article/91f5ed2ea2764533861ff05194d6953f2017-05-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01977-16https://doaj.org/toc/2150-7511ABSTRACT Perturbation of cellular processes is a prevailing approach to understanding biology. To better understand the complicated biology that defines bacterial shape, a sensitive, high-content platform was developed to detect multiple morphological defect phenotypes using microscopy. We examined morphological phenotypes across the Escherichia coli K-12 deletion (Keio) collection at the mid-exponential growth phase, revealing 111 deletions perturbing shape. Interestingly, 64% of these were uncharacterized mutants, illustrating the complex nature of shape maintenance and regulation in bacteria. To understand the roles these genes play in defining morphology, 53 mutants with knockouts resulting in abnormal cell shape were crossed with the Keio collection in high throughput, generating 1,373 synthetic lethal interactions across 1.7 million double deletion mutants. This analysis yielded a highly populated interaction network spanning and linking multiple phenotypes, with a preponderance of interactions involved in transport, oxidation-reduction, and metabolic processes. IMPORTANCE Genetic perturbations of cellular functions are a prevailing approach to understanding cell systems, which are increasingly being practiced in very high throughput. Here, we report a high-content microscopy platform tailored to bacteria, which probes the impact of genetic mutation on cell morphology. This has particular utility in revealing elusive and subtle morphological phenotypes associated with blocks in nonessential cellular functions. We report 111 nonessential mutations impacting E. coli morphology, with nearly half of those genes being poorly annotated or uncharacterized. Further, these genes appear to be tightly linked to transport or redox processes within the cell. The screening platform is simple and low cost and is broadly applicable to any bacterial genomic library or chemical collection. Indeed, this is a powerful tool in understanding the biology behind bacterial shape.Shawn FrenchJean-Philippe CôtéJonathan M. StokesRay TruantEric D. BrownAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 8, Iss 2 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Shawn French Jean-Philippe Côté Jonathan M. Stokes Ray Truant Eric D. Brown Bacteria Getting into Shape: Genetic Determinants of <italic toggle="yes">E. coli</italic> Morphology
description	ABSTRACT Perturbation of cellular processes is a prevailing approach to understanding biology. To better understand the complicated biology that defines bacterial shape, a sensitive, high-content platform was developed to detect multiple morphological defect phenotypes using microscopy. We examined morphological phenotypes across the Escherichia coli K-12 deletion (Keio) collection at the mid-exponential growth phase, revealing 111 deletions perturbing shape. Interestingly, 64% of these were uncharacterized mutants, illustrating the complex nature of shape maintenance and regulation in bacteria. To understand the roles these genes play in defining morphology, 53 mutants with knockouts resulting in abnormal cell shape were crossed with the Keio collection in high throughput, generating 1,373 synthetic lethal interactions across 1.7 million double deletion mutants. This analysis yielded a highly populated interaction network spanning and linking multiple phenotypes, with a preponderance of interactions involved in transport, oxidation-reduction, and metabolic processes. IMPORTANCE Genetic perturbations of cellular functions are a prevailing approach to understanding cell systems, which are increasingly being practiced in very high throughput. Here, we report a high-content microscopy platform tailored to bacteria, which probes the impact of genetic mutation on cell morphology. This has particular utility in revealing elusive and subtle morphological phenotypes associated with blocks in nonessential cellular functions. We report 111 nonessential mutations impacting E. coli morphology, with nearly half of those genes being poorly annotated or uncharacterized. Further, these genes appear to be tightly linked to transport or redox processes within the cell. The screening platform is simple and low cost and is broadly applicable to any bacterial genomic library or chemical collection. Indeed, this is a powerful tool in understanding the biology behind bacterial shape.
format	article
author	Shawn French Jean-Philippe Côté Jonathan M. Stokes Ray Truant Eric D. Brown
author_facet	Shawn French Jean-Philippe Côté Jonathan M. Stokes Ray Truant Eric D. Brown
author_sort	Shawn French
title	Bacteria Getting into Shape: Genetic Determinants of <italic toggle="yes">E. coli</italic> Morphology
title_short	Bacteria Getting into Shape: Genetic Determinants of <italic toggle="yes">E. coli</italic> Morphology
title_full	Bacteria Getting into Shape: Genetic Determinants of <italic toggle="yes">E. coli</italic> Morphology
title_fullStr	Bacteria Getting into Shape: Genetic Determinants of <italic toggle="yes">E. coli</italic> Morphology
title_full_unstemmed	Bacteria Getting into Shape: Genetic Determinants of <italic toggle="yes">E. coli</italic> Morphology
title_sort	bacteria getting into shape: genetic determinants of <italic toggle="yes">e. coli</italic> morphology
publisher	American Society for Microbiology
publishDate	2017
url	https://doaj.org/article/91f5ed2ea2764533861ff05194d6953f
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Bacteria Getting into Shape: Genetic Determinants of <italic toggle="yes">E. coli</italic> Morphology

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