Genome-Wide Identification of <named-content content-type="genus-species">Acinetobacter baumannii</named-content> Genes Necessary for Persistence in the Lung

ABSTRACT Acinetobacter baumannii is a Gram-negative bacterium that causes diseases such as pneumonia, bacteremia, and soft tissue infections in hospitalized patients. Relatively little is known about how A. baumannii causes these infections. Thus, we used insertion sequencing (INSeq), a combination...

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Autores principales:	Nengding Wang, Egon A. Ozer, Mark J. Mandel, Alan R. Hauser
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Lenguaje:	EN
Publicado:	American Society for Microbiology 2014
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/9cabbfad40864b4393adae7654985ac3
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spelling	oai:doaj.org-article:9cabbfad40864b4393adae7654985ac32021-11-15T15:47:38ZGenome-Wide Identification of <named-content content-type="genus-species">Acinetobacter baumannii</named-content> Genes Necessary for Persistence in the Lung10.1128/mBio.01163-142150-7511https://doaj.org/article/9cabbfad40864b4393adae7654985ac32014-07-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01163-14https://doaj.org/toc/2150-7511ABSTRACT Acinetobacter baumannii is a Gram-negative bacterium that causes diseases such as pneumonia, bacteremia, and soft tissue infections in hospitalized patients. Relatively little is known about how A. baumannii causes these infections. Thus, we used insertion sequencing (INSeq), a combination of transposon mutagenesis and massively parallel next-generation sequencing, to identify novel virulence factors of A. baumannii. To this end, we generated a random transposon mutant library containing 150,000 unique insertions in A. baumannii strain ATCC 17978. The INSeq analysis identified 453 genes required for growth in rich medium. The library was then used in a murine pneumonia model, and the relative levels of abundance of mutants before and after selection in the mouse were compared. When genes required for growth in rich medium were removed from the analysis, 157 genes were identified as necessary for persistence in the mouse lung. Several of these encode known virulence factors of A. baumannii, such as OmpA and ZnuB, which validated our approach. A large number of the genes identified were predicted to be involved in amino acid and nucleotide metabolism and transport. Other genes were predicted to encode an integration host factor, a transmembrane lipoprotein, and proteins involved in stress response and efflux pumps. Very few genes, when disrupted, resulted in an increase in A. baumannii numbers during host infection. The INSeq approach identified a number of novel virulence determinants of A. baumannii, which are candidate targets for therapeutic interventions. IMPORTANCE A. baumannii has emerged as a frequent cause of serious infections in hospitals and community settings. Due to increasing antibiotic resistance, alternative approaches, such as antivirulence strategies, are desperately needed to fight A. baumannii infections. Thorough knowledge of A. baumannii pathogenicity is essential for such approaches but is currently lacking. With the increasingly widespread use of massively parallel sequencing, a class of techniques known as transposon insertion sequencing has been developed to perform comprehensive virulence screens of bacterial genomes in vivo. We have applied one of these approaches (INSeq) to uncover novel virulence factors in A. baumannii. We identified several such factors, including those predicted to encode amino acid and nucleotide metabolism proteins, an integration host factor protein, stress response factors, and efflux pumps. These results greatly expand the number of A. baumannii virulence factors and uncover potential targets for antivirulence treatments.Nengding WangEgon A. OzerMark J. MandelAlan R. HauserAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 5, Iss 3 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Nengding Wang Egon A. Ozer Mark J. Mandel Alan R. Hauser Genome-Wide Identification of <named-content content-type="genus-species">Acinetobacter baumannii</named-content> Genes Necessary for Persistence in the Lung
description	ABSTRACT Acinetobacter baumannii is a Gram-negative bacterium that causes diseases such as pneumonia, bacteremia, and soft tissue infections in hospitalized patients. Relatively little is known about how A. baumannii causes these infections. Thus, we used insertion sequencing (INSeq), a combination of transposon mutagenesis and massively parallel next-generation sequencing, to identify novel virulence factors of A. baumannii. To this end, we generated a random transposon mutant library containing 150,000 unique insertions in A. baumannii strain ATCC 17978. The INSeq analysis identified 453 genes required for growth in rich medium. The library was then used in a murine pneumonia model, and the relative levels of abundance of mutants before and after selection in the mouse were compared. When genes required for growth in rich medium were removed from the analysis, 157 genes were identified as necessary for persistence in the mouse lung. Several of these encode known virulence factors of A. baumannii, such as OmpA and ZnuB, which validated our approach. A large number of the genes identified were predicted to be involved in amino acid and nucleotide metabolism and transport. Other genes were predicted to encode an integration host factor, a transmembrane lipoprotein, and proteins involved in stress response and efflux pumps. Very few genes, when disrupted, resulted in an increase in A. baumannii numbers during host infection. The INSeq approach identified a number of novel virulence determinants of A. baumannii, which are candidate targets for therapeutic interventions. IMPORTANCE A. baumannii has emerged as a frequent cause of serious infections in hospitals and community settings. Due to increasing antibiotic resistance, alternative approaches, such as antivirulence strategies, are desperately needed to fight A. baumannii infections. Thorough knowledge of A. baumannii pathogenicity is essential for such approaches but is currently lacking. With the increasingly widespread use of massively parallel sequencing, a class of techniques known as transposon insertion sequencing has been developed to perform comprehensive virulence screens of bacterial genomes in vivo. We have applied one of these approaches (INSeq) to uncover novel virulence factors in A. baumannii. We identified several such factors, including those predicted to encode amino acid and nucleotide metabolism proteins, an integration host factor protein, stress response factors, and efflux pumps. These results greatly expand the number of A. baumannii virulence factors and uncover potential targets for antivirulence treatments.
format	article
author	Nengding Wang Egon A. Ozer Mark J. Mandel Alan R. Hauser
author_facet	Nengding Wang Egon A. Ozer Mark J. Mandel Alan R. Hauser
author_sort	Nengding Wang
title	Genome-Wide Identification of <named-content content-type="genus-species">Acinetobacter baumannii</named-content> Genes Necessary for Persistence in the Lung
title_short	Genome-Wide Identification of <named-content content-type="genus-species">Acinetobacter baumannii</named-content> Genes Necessary for Persistence in the Lung
title_full	Genome-Wide Identification of <named-content content-type="genus-species">Acinetobacter baumannii</named-content> Genes Necessary for Persistence in the Lung
title_fullStr	Genome-Wide Identification of <named-content content-type="genus-species">Acinetobacter baumannii</named-content> Genes Necessary for Persistence in the Lung
title_full_unstemmed	Genome-Wide Identification of <named-content content-type="genus-species">Acinetobacter baumannii</named-content> Genes Necessary for Persistence in the Lung
title_sort	genome-wide identification of <named-content content-type="genus-species">acinetobacter baumannii</named-content> genes necessary for persistence in the lung
publisher	American Society for Microbiology
publishDate	2014
url	https://doaj.org/article/9cabbfad40864b4393adae7654985ac3
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Genome-Wide Identification of <named-content content-type="genus-species">Acinetobacter baumannii</named-content> Genes Necessary for Persistence in the Lung

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