Stochastic Variation in Expression of the Tricarboxylic Acid Cycle Produces Persister Cells

ABSTRACT Chronic bacterial infections are difficult to eradicate, though they are caused primarily by drug-susceptible pathogens. Antibiotic-tolerant persisters largely account for this paradox. In spite of their significance in the recalcitrance of chronic infections, the mechanism of persister for...

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Autores principales:	Eliza A. Zalis, Austin S. Nuxoll, Sylvie Manuse, Geremy Clair, Lauren C. Radlinski, Brian P. Conlon, Joshua Adkins, Kim Lewis
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	Staphylococcus aureus bioenergetics heterologous gene expression metabolism persistence tolerance Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/7ed88a0dd9c64c25b08618a700f515f2
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spelling	oai:doaj.org-article:7ed88a0dd9c64c25b08618a700f515f22021-11-15T15:59:41ZStochastic Variation in Expression of the Tricarboxylic Acid Cycle Produces Persister Cells10.1128/mBio.01930-192150-7511https://doaj.org/article/7ed88a0dd9c64c25b08618a700f515f22019-10-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01930-19https://doaj.org/toc/2150-7511ABSTRACT Chronic bacterial infections are difficult to eradicate, though they are caused primarily by drug-susceptible pathogens. Antibiotic-tolerant persisters largely account for this paradox. In spite of their significance in the recalcitrance of chronic infections, the mechanism of persister formation is poorly understood. We previously reported that a decrease in ATP levels leads to drug tolerance in Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus. We reasoned that stochastic fluctuation in the expression of tricarboxylic acid (TCA) cycle enzymes can produce cells with low energy levels. S. aureus knockouts in glutamate dehydrogenase, 2-oxoketoglutarate dehydrogenase, succinyl coenzyme A (CoA) synthetase, and fumarase have low ATP levels and exhibit increased tolerance of fluoroquinolone, aminoglycoside, and β-lactam antibiotics. Fluorescence-activated cell sorter (FACS) analysis of TCA genes shows a broad Gaussian distribution in a population, with differences of over 3 orders of magnitude in the levels of expression between individual cells. Sorted cells with low levels of TCA enzyme expression have an increased tolerance of antibiotic treatment. These findings suggest that fluctuations in the levels of expression of energy-generating components serve as a mechanism of persister formation. IMPORTANCE Persister cells are rare phenotypic variants that are able to survive antibiotic treatment. Unlike resistant bacteria, which have specific mechanisms to prevent antibiotics from binding to their targets, persisters evade antibiotic killing by entering a tolerant nongrowing state. Persisters have been implicated in chronic infections in multiple species, and growing evidence suggests that persister cells are responsible for many cases of antibiotic treatment failure. New antibiotic treatment strategies aim to kill tolerant persister cells more effectively, but the mechanism of tolerance has remained unclear until now.Eliza A. ZalisAustin S. NuxollSylvie ManuseGeremy ClairLauren C. RadlinskiBrian P. ConlonJoshua AdkinsKim LewisAmerican Society for MicrobiologyarticleStaphylococcus aureusbioenergeticsheterologous gene expressionmetabolismpersistencetoleranceMicrobiologyQR1-502ENmBio, Vol 10, Iss 5 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	Staphylococcus aureus bioenergetics heterologous gene expression metabolism persistence tolerance Microbiology QR1-502
spellingShingle	Staphylococcus aureus bioenergetics heterologous gene expression metabolism persistence tolerance Microbiology QR1-502 Eliza A. Zalis Austin S. Nuxoll Sylvie Manuse Geremy Clair Lauren C. Radlinski Brian P. Conlon Joshua Adkins Kim Lewis Stochastic Variation in Expression of the Tricarboxylic Acid Cycle Produces Persister Cells
description	ABSTRACT Chronic bacterial infections are difficult to eradicate, though they are caused primarily by drug-susceptible pathogens. Antibiotic-tolerant persisters largely account for this paradox. In spite of their significance in the recalcitrance of chronic infections, the mechanism of persister formation is poorly understood. We previously reported that a decrease in ATP levels leads to drug tolerance in Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus. We reasoned that stochastic fluctuation in the expression of tricarboxylic acid (TCA) cycle enzymes can produce cells with low energy levels. S. aureus knockouts in glutamate dehydrogenase, 2-oxoketoglutarate dehydrogenase, succinyl coenzyme A (CoA) synthetase, and fumarase have low ATP levels and exhibit increased tolerance of fluoroquinolone, aminoglycoside, and β-lactam antibiotics. Fluorescence-activated cell sorter (FACS) analysis of TCA genes shows a broad Gaussian distribution in a population, with differences of over 3 orders of magnitude in the levels of expression between individual cells. Sorted cells with low levels of TCA enzyme expression have an increased tolerance of antibiotic treatment. These findings suggest that fluctuations in the levels of expression of energy-generating components serve as a mechanism of persister formation. IMPORTANCE Persister cells are rare phenotypic variants that are able to survive antibiotic treatment. Unlike resistant bacteria, which have specific mechanisms to prevent antibiotics from binding to their targets, persisters evade antibiotic killing by entering a tolerant nongrowing state. Persisters have been implicated in chronic infections in multiple species, and growing evidence suggests that persister cells are responsible for many cases of antibiotic treatment failure. New antibiotic treatment strategies aim to kill tolerant persister cells more effectively, but the mechanism of tolerance has remained unclear until now.
format	article
author	Eliza A. Zalis Austin S. Nuxoll Sylvie Manuse Geremy Clair Lauren C. Radlinski Brian P. Conlon Joshua Adkins Kim Lewis
author_facet	Eliza A. Zalis Austin S. Nuxoll Sylvie Manuse Geremy Clair Lauren C. Radlinski Brian P. Conlon Joshua Adkins Kim Lewis
author_sort	Eliza A. Zalis
title	Stochastic Variation in Expression of the Tricarboxylic Acid Cycle Produces Persister Cells
title_short	Stochastic Variation in Expression of the Tricarboxylic Acid Cycle Produces Persister Cells
title_full	Stochastic Variation in Expression of the Tricarboxylic Acid Cycle Produces Persister Cells
title_fullStr	Stochastic Variation in Expression of the Tricarboxylic Acid Cycle Produces Persister Cells
title_full_unstemmed	Stochastic Variation in Expression of the Tricarboxylic Acid Cycle Produces Persister Cells
title_sort	stochastic variation in expression of the tricarboxylic acid cycle produces persister cells
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/7ed88a0dd9c64c25b08618a700f515f2
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Stochastic Variation in Expression of the Tricarboxylic Acid Cycle Produces Persister Cells

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