A Shift in Central Metabolism Accompanies Virulence Activation in <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content>

ABSTRACT The availability of energy has significant impact on cell physiology. However, the role of cellular metabolism in bacterial pathogenesis is not understood. We investigated the dynamics of central metabolism during virulence induction by surface sensing and quorum sensing in early-stage biof...

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Autores principales:	Kumar Perinbam, Jenu V. Chacko, Anerudh Kannan, Michelle A. Digman, Albert Siryaporn
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	FLIM PilY1 antivirulence central metabolism fluorescence lifetime imaging microscopy quorum sensing Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/7063abd4513c44378e6a216c3a669c9b
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spelling	oai:doaj.org-article:7063abd4513c44378e6a216c3a669c9b2021-11-15T15:57:02ZA Shift in Central Metabolism Accompanies Virulence Activation in <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content>10.1128/mBio.02730-182150-7511https://doaj.org/article/7063abd4513c44378e6a216c3a669c9b2020-04-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.02730-18https://doaj.org/toc/2150-7511ABSTRACT The availability of energy has significant impact on cell physiology. However, the role of cellular metabolism in bacterial pathogenesis is not understood. We investigated the dynamics of central metabolism during virulence induction by surface sensing and quorum sensing in early-stage biofilms of the multidrug-resistant bacterium Pseudomonas aeruginosa. We established a metabolic profile for P. aeruginosa using fluorescence lifetime imaging microscopy (FLIM), which reports the activity of NADH in live cells. We identified a critical growth transition period during which virulence is activated. We performed FLIM measurements and direct measurements of NADH and NAD+ concentrations during this period. Here, planktonic (low-virulence) and surface-attached (virulence-activated) populations diverged into distinct metabolic states, with the surface-attached population exhibiting FLIM lifetimes that were associated with lower levels of enzyme-bound NADH and decreasing total NAD(H) production. We inhibited virulence by perturbing central metabolism using citrate and pyruvate, which further decreased the enzyme-bound NADH fraction and total NAD(H) production and suggested the involvement of the glyoxylate pathway in virulence activation in surface-attached populations. In addition, we induced virulence at an earlier time using the electron transport chain oxidase inhibitor antimycin A. Our results demonstrate the use of FLIM to noninvasively measure NADH dynamics in biofilms and suggest a model in which a metabolic rearrangement accompanies the virulence activation period. IMPORTANCE The rise of antibiotic resistance requires the development of new strategies to combat bacterial infection and pathogenesis. A major direction has been the development of drugs that broadly target virulence. However, few targets have been identified due to the species-specific nature of many virulence regulators. The lack of a virulence regulator that is conserved across species has presented a further challenge to the development of therapeutics. Here, we identify that NADH activity has an important role in the induction of virulence in the pathogen P. aeruginosa. This finding, coupled with the ubiquity of NADH in bacterial pathogens, opens up the possibility of targeting enzymes that process NADH as a potential broad antivirulence approach.Kumar PerinbamJenu V. ChackoAnerudh KannanMichelle A. DigmanAlbert SiryapornAmerican Society for MicrobiologyarticleFLIMPilY1antivirulencecentral metabolismfluorescence lifetime imaging microscopyquorum sensingMicrobiologyQR1-502ENmBio, Vol 11, Iss 2 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	FLIM PilY1 antivirulence central metabolism fluorescence lifetime imaging microscopy quorum sensing Microbiology QR1-502
spellingShingle	FLIM PilY1 antivirulence central metabolism fluorescence lifetime imaging microscopy quorum sensing Microbiology QR1-502 Kumar Perinbam Jenu V. Chacko Anerudh Kannan Michelle A. Digman Albert Siryaporn A Shift in Central Metabolism Accompanies Virulence Activation in <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content>
description	ABSTRACT The availability of energy has significant impact on cell physiology. However, the role of cellular metabolism in bacterial pathogenesis is not understood. We investigated the dynamics of central metabolism during virulence induction by surface sensing and quorum sensing in early-stage biofilms of the multidrug-resistant bacterium Pseudomonas aeruginosa. We established a metabolic profile for P. aeruginosa using fluorescence lifetime imaging microscopy (FLIM), which reports the activity of NADH in live cells. We identified a critical growth transition period during which virulence is activated. We performed FLIM measurements and direct measurements of NADH and NAD+ concentrations during this period. Here, planktonic (low-virulence) and surface-attached (virulence-activated) populations diverged into distinct metabolic states, with the surface-attached population exhibiting FLIM lifetimes that were associated with lower levels of enzyme-bound NADH and decreasing total NAD(H) production. We inhibited virulence by perturbing central metabolism using citrate and pyruvate, which further decreased the enzyme-bound NADH fraction and total NAD(H) production and suggested the involvement of the glyoxylate pathway in virulence activation in surface-attached populations. In addition, we induced virulence at an earlier time using the electron transport chain oxidase inhibitor antimycin A. Our results demonstrate the use of FLIM to noninvasively measure NADH dynamics in biofilms and suggest a model in which a metabolic rearrangement accompanies the virulence activation period. IMPORTANCE The rise of antibiotic resistance requires the development of new strategies to combat bacterial infection and pathogenesis. A major direction has been the development of drugs that broadly target virulence. However, few targets have been identified due to the species-specific nature of many virulence regulators. The lack of a virulence regulator that is conserved across species has presented a further challenge to the development of therapeutics. Here, we identify that NADH activity has an important role in the induction of virulence in the pathogen P. aeruginosa. This finding, coupled with the ubiquity of NADH in bacterial pathogens, opens up the possibility of targeting enzymes that process NADH as a potential broad antivirulence approach.
format	article
author	Kumar Perinbam Jenu V. Chacko Anerudh Kannan Michelle A. Digman Albert Siryaporn
author_facet	Kumar Perinbam Jenu V. Chacko Anerudh Kannan Michelle A. Digman Albert Siryaporn
author_sort	Kumar Perinbam
title	A Shift in Central Metabolism Accompanies Virulence Activation in <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content>
title_short	A Shift in Central Metabolism Accompanies Virulence Activation in <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content>
title_full	A Shift in Central Metabolism Accompanies Virulence Activation in <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content>
title_fullStr	A Shift in Central Metabolism Accompanies Virulence Activation in <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content>
title_full_unstemmed	A Shift in Central Metabolism Accompanies Virulence Activation in <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content>
title_sort	shift in central metabolism accompanies virulence activation in <named-content content-type="genus-species">pseudomonas aeruginosa</named-content>
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/7063abd4513c44378e6a216c3a669c9b
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A Shift in Central Metabolism Accompanies Virulence Activation in <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content>

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