A High-Content, Phenotypic Screen Identifies Fluorouridine as an Inhibitor of Pyoverdine Biosynthesis and <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content> Virulence

ABSTRACT Pseudomonas aeruginosa is an opportunistic pathogen that causes severe health problems. Despite intensive investigation, many aspects of microbial virulence remain poorly understood. We used a high-throughput, high-content, whole-organism, phenotypic screen to identify small molecules that...

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Autores principales:	Daniel R. Kirienko, Alexey V. Revtovich, Natalia V. Kirienko
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2016
Materias:	5-fluorouracil 5-fluorouridine Caenorhabditis elegans Pseudomonas aeruginosa pyoverdine high-throughput screening Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/8233a6fa30344edb9669d20c4b479792
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spelling	oai:doaj.org-article:8233a6fa30344edb9669d20c4b4797922021-11-15T15:21:14ZA High-Content, Phenotypic Screen Identifies Fluorouridine as an Inhibitor of Pyoverdine Biosynthesis and <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content> Virulence10.1128/mSphere.00217-162379-5042https://doaj.org/article/8233a6fa30344edb9669d20c4b4797922016-08-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00217-16https://doaj.org/toc/2379-5042ABSTRACT Pseudomonas aeruginosa is an opportunistic pathogen that causes severe health problems. Despite intensive investigation, many aspects of microbial virulence remain poorly understood. We used a high-throughput, high-content, whole-organism, phenotypic screen to identify small molecules that inhibit P. aeruginosa virulence in Caenorhabditis elegans. Approximately half of the hits were known antimicrobials. A large number of hits were nonantimicrobial bioactive compounds, including the cancer chemotherapeutic 5-fluorouracil. We determined that 5-fluorouracil both transiently inhibits bacterial growth and reduces pyoverdine biosynthesis. Pyoverdine is a siderophore that regulates the expression of several virulence determinants and is critical for pathogenesis in mammals. We show that 5-fluorouridine, a downstream metabolite of 5-fluorouracil, is responsible for inhibiting pyoverdine biosynthesis. We also show that 5-fluorouridine, in contrast to 5-fluorouracil, is a genuine antivirulence compound, with no bacteriostatic or bactericidal activity. To our knowledge, this is the first report utilizing a whole-organism screen to identify novel compounds with antivirulent properties effective against P. aeruginosa. IMPORTANCE Despite intense research effort from scientists and the advent of the molecular age of biomedical research, many of the mechanisms that underlie pathogenesis are still understood poorly, if at all. The opportunistic human pathogen Pseudomonas aeruginosa causes a variety of soft tissue infections and is responsible for over 50,000 hospital-acquired infections per year. In addition, P. aeruginosa exhibits a striking degree of innate and acquired antimicrobial resistance, complicating treatment. It is increasingly important to understand P. aeruginosa virulence. In an effort to gain this information in an unbiased fashion, we used a high-throughput phenotypic screen to identify small molecules that disrupted bacterial pathogenesis and increased host survival using the model nematode Caenorhabditis elegans. This method led to the unexpected discovery that addition of a modified nucleotide, 5-fluorouridine, disrupted bacterial RNA metabolism and inhibited synthesis of pyoverdine, a critical toxin. Our results demonstrate that this compound specifically functions as an antivirulent.Daniel R. KirienkoAlexey V. RevtovichNatalia V. KirienkoAmerican Society for Microbiologyarticle5-fluorouracil5-fluorouridineCaenorhabditis elegansPseudomonas aeruginosapyoverdinehigh-throughput screeningMicrobiologyQR1-502ENmSphere, Vol 1, Iss 4 (2016)
institution	DOAJ
collection	DOAJ
language	EN
topic	5-fluorouracil 5-fluorouridine Caenorhabditis elegans Pseudomonas aeruginosa pyoverdine high-throughput screening Microbiology QR1-502
spellingShingle	5-fluorouracil 5-fluorouridine Caenorhabditis elegans Pseudomonas aeruginosa pyoverdine high-throughput screening Microbiology QR1-502 Daniel R. Kirienko Alexey V. Revtovich Natalia V. Kirienko A High-Content, Phenotypic Screen Identifies Fluorouridine as an Inhibitor of Pyoverdine Biosynthesis and <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content> Virulence
description	ABSTRACT Pseudomonas aeruginosa is an opportunistic pathogen that causes severe health problems. Despite intensive investigation, many aspects of microbial virulence remain poorly understood. We used a high-throughput, high-content, whole-organism, phenotypic screen to identify small molecules that inhibit P. aeruginosa virulence in Caenorhabditis elegans. Approximately half of the hits were known antimicrobials. A large number of hits were nonantimicrobial bioactive compounds, including the cancer chemotherapeutic 5-fluorouracil. We determined that 5-fluorouracil both transiently inhibits bacterial growth and reduces pyoverdine biosynthesis. Pyoverdine is a siderophore that regulates the expression of several virulence determinants and is critical for pathogenesis in mammals. We show that 5-fluorouridine, a downstream metabolite of 5-fluorouracil, is responsible for inhibiting pyoverdine biosynthesis. We also show that 5-fluorouridine, in contrast to 5-fluorouracil, is a genuine antivirulence compound, with no bacteriostatic or bactericidal activity. To our knowledge, this is the first report utilizing a whole-organism screen to identify novel compounds with antivirulent properties effective against P. aeruginosa. IMPORTANCE Despite intense research effort from scientists and the advent of the molecular age of biomedical research, many of the mechanisms that underlie pathogenesis are still understood poorly, if at all. The opportunistic human pathogen Pseudomonas aeruginosa causes a variety of soft tissue infections and is responsible for over 50,000 hospital-acquired infections per year. In addition, P. aeruginosa exhibits a striking degree of innate and acquired antimicrobial resistance, complicating treatment. It is increasingly important to understand P. aeruginosa virulence. In an effort to gain this information in an unbiased fashion, we used a high-throughput phenotypic screen to identify small molecules that disrupted bacterial pathogenesis and increased host survival using the model nematode Caenorhabditis elegans. This method led to the unexpected discovery that addition of a modified nucleotide, 5-fluorouridine, disrupted bacterial RNA metabolism and inhibited synthesis of pyoverdine, a critical toxin. Our results demonstrate that this compound specifically functions as an antivirulent.
format	article
author	Daniel R. Kirienko Alexey V. Revtovich Natalia V. Kirienko
author_facet	Daniel R. Kirienko Alexey V. Revtovich Natalia V. Kirienko
author_sort	Daniel R. Kirienko
title	A High-Content, Phenotypic Screen Identifies Fluorouridine as an Inhibitor of Pyoverdine Biosynthesis and <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content> Virulence
title_short	A High-Content, Phenotypic Screen Identifies Fluorouridine as an Inhibitor of Pyoverdine Biosynthesis and <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content> Virulence
title_full	A High-Content, Phenotypic Screen Identifies Fluorouridine as an Inhibitor of Pyoverdine Biosynthesis and <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content> Virulence
title_fullStr	A High-Content, Phenotypic Screen Identifies Fluorouridine as an Inhibitor of Pyoverdine Biosynthesis and <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content> Virulence
title_full_unstemmed	A High-Content, Phenotypic Screen Identifies Fluorouridine as an Inhibitor of Pyoverdine Biosynthesis and <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content> Virulence
title_sort	high-content, phenotypic screen identifies fluorouridine as an inhibitor of pyoverdine biosynthesis and <named-content content-type="genus-species">pseudomonas aeruginosa</named-content> virulence
publisher	American Society for Microbiology
publishDate	2016
url	https://doaj.org/article/8233a6fa30344edb9669d20c4b479792
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A High-Content, Phenotypic Screen Identifies Fluorouridine as an Inhibitor of Pyoverdine Biosynthesis and <named-content content-type="genus-species">Pseudomonas aeruginosa</named-content> Virulence

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