Genome-Wide Mutant Fitness Profiling Identifies Nutritional Requirements for Optimal Growth of <named-content content-type="genus-species">Yersinia pestis</named-content> in Deep Tissue

ABSTRACT Rapid growth in deep tissue is essential to the high virulence of Yersinia pestis, causative agent of plague. To better understand the mechanisms underlying this unusual ability, we used transposon mutagenesis and high-throughput sequencing (Tn-seq) to systematically probe the Y. pestis gen...

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Autores principales:	Samantha G. Palace, Megan K. Proulx, Shan Lu, Richard E. Baker, Jon D. Goguen
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2014
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/669f11aa0a76439dbf5546c1a1894602
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spelling	oai:doaj.org-article:669f11aa0a76439dbf5546c1a18946022021-11-15T15:47:21ZGenome-Wide Mutant Fitness Profiling Identifies Nutritional Requirements for Optimal Growth of <named-content content-type="genus-species">Yersinia pestis</named-content> in Deep Tissue10.1128/mBio.01385-142150-7511https://doaj.org/article/669f11aa0a76439dbf5546c1a18946022014-08-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01385-14https://doaj.org/toc/2150-7511ABSTRACT Rapid growth in deep tissue is essential to the high virulence of Yersinia pestis, causative agent of plague. To better understand the mechanisms underlying this unusual ability, we used transposon mutagenesis and high-throughput sequencing (Tn-seq) to systematically probe the Y. pestis genome for elements contributing to fitness during infection. More than a million independent insertion mutants representing nearly 200,000 unique genotypes were generated in fully virulent Y. pestis. Each mutant in the library was assayed for its ability to proliferate in vitro on rich medium and in mice following intravenous injection. Virtually all genes previously established to contribute to virulence following intravenous infection showed significant fitness defects, with the exception of genes for yersiniabactin biosynthesis, which were masked by strong intercellular complementation effects. We also identified more than 30 genes with roles in nutrient acquisition and metabolism as experiencing strong selection during infection. Many of these genes had not previously been implicated in Y. pestis virulence. We further examined the fitness defects of strains carrying mutations in two such genes—encoding a branched-chain amino acid importer (brnQ) and a glucose importer (ptsG)—both in vivo and in a novel defined synthetic growth medium with nutrient concentrations matching those in serum. Our findings suggest that diverse nutrient limitations in deep tissue play a more important role in controlling bacterial infection than has heretofore been appreciated. Because much is known about Y. pestis pathogenesis, this study also serves as a test case that assesses the ability of Tn-seq to detect virulence genes. IMPORTANCE Our understanding of the functions required by bacteria to grow in deep tissues is limited, in part because most growth studies of pathogenic bacteria are conducted on laboratory media that do not reflect conditions prevailing in infected animal tissues. Improving our knowledge of this aspect of bacterial biology is important as a potential pathway to the development of novel therapeutics. Yersinia pestis, the plague bacterium, is highly virulent due to its rapid dissemination and growth in deep tissues, making it a good model for discovering bacterial adaptations that promote rapid growth during infection. Using Tn-seq, a genome-wide fitness profiling technique, we identified several functions required for fitness of Y. pestis in vivo that were not previously known to be important. Most of these functions are needed to acquire or synthesize nutrients. Interference with these critical nutrient acquisition pathways may be an effective strategy for designing novel antibiotics and vaccines.Samantha G. PalaceMegan K. ProulxShan LuRichard E. BakerJon D. GoguenAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 5, Iss 4 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Samantha G. Palace Megan K. Proulx Shan Lu Richard E. Baker Jon D. Goguen Genome-Wide Mutant Fitness Profiling Identifies Nutritional Requirements for Optimal Growth of <named-content content-type="genus-species">Yersinia pestis</named-content> in Deep Tissue
description	ABSTRACT Rapid growth in deep tissue is essential to the high virulence of Yersinia pestis, causative agent of plague. To better understand the mechanisms underlying this unusual ability, we used transposon mutagenesis and high-throughput sequencing (Tn-seq) to systematically probe the Y. pestis genome for elements contributing to fitness during infection. More than a million independent insertion mutants representing nearly 200,000 unique genotypes were generated in fully virulent Y. pestis. Each mutant in the library was assayed for its ability to proliferate in vitro on rich medium and in mice following intravenous injection. Virtually all genes previously established to contribute to virulence following intravenous infection showed significant fitness defects, with the exception of genes for yersiniabactin biosynthesis, which were masked by strong intercellular complementation effects. We also identified more than 30 genes with roles in nutrient acquisition and metabolism as experiencing strong selection during infection. Many of these genes had not previously been implicated in Y. pestis virulence. We further examined the fitness defects of strains carrying mutations in two such genes—encoding a branched-chain amino acid importer (brnQ) and a glucose importer (ptsG)—both in vivo and in a novel defined synthetic growth medium with nutrient concentrations matching those in serum. Our findings suggest that diverse nutrient limitations in deep tissue play a more important role in controlling bacterial infection than has heretofore been appreciated. Because much is known about Y. pestis pathogenesis, this study also serves as a test case that assesses the ability of Tn-seq to detect virulence genes. IMPORTANCE Our understanding of the functions required by bacteria to grow in deep tissues is limited, in part because most growth studies of pathogenic bacteria are conducted on laboratory media that do not reflect conditions prevailing in infected animal tissues. Improving our knowledge of this aspect of bacterial biology is important as a potential pathway to the development of novel therapeutics. Yersinia pestis, the plague bacterium, is highly virulent due to its rapid dissemination and growth in deep tissues, making it a good model for discovering bacterial adaptations that promote rapid growth during infection. Using Tn-seq, a genome-wide fitness profiling technique, we identified several functions required for fitness of Y. pestis in vivo that were not previously known to be important. Most of these functions are needed to acquire or synthesize nutrients. Interference with these critical nutrient acquisition pathways may be an effective strategy for designing novel antibiotics and vaccines.
format	article
author	Samantha G. Palace Megan K. Proulx Shan Lu Richard E. Baker Jon D. Goguen
author_facet	Samantha G. Palace Megan K. Proulx Shan Lu Richard E. Baker Jon D. Goguen
author_sort	Samantha G. Palace
title	Genome-Wide Mutant Fitness Profiling Identifies Nutritional Requirements for Optimal Growth of <named-content content-type="genus-species">Yersinia pestis</named-content> in Deep Tissue
title_short	Genome-Wide Mutant Fitness Profiling Identifies Nutritional Requirements for Optimal Growth of <named-content content-type="genus-species">Yersinia pestis</named-content> in Deep Tissue
title_full	Genome-Wide Mutant Fitness Profiling Identifies Nutritional Requirements for Optimal Growth of <named-content content-type="genus-species">Yersinia pestis</named-content> in Deep Tissue
title_fullStr	Genome-Wide Mutant Fitness Profiling Identifies Nutritional Requirements for Optimal Growth of <named-content content-type="genus-species">Yersinia pestis</named-content> in Deep Tissue
title_full_unstemmed	Genome-Wide Mutant Fitness Profiling Identifies Nutritional Requirements for Optimal Growth of <named-content content-type="genus-species">Yersinia pestis</named-content> in Deep Tissue
title_sort	genome-wide mutant fitness profiling identifies nutritional requirements for optimal growth of <named-content content-type="genus-species">yersinia pestis</named-content> in deep tissue
publisher	American Society for Microbiology
publishDate	2014
url	https://doaj.org/article/669f11aa0a76439dbf5546c1a1894602
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Genome-Wide Mutant Fitness Profiling Identifies Nutritional Requirements for Optimal Growth of <named-content content-type="genus-species">Yersinia pestis</named-content> in Deep Tissue

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