High-Throughput Analysis of Gene Function in the Bacterial Predator <named-content content-type="genus-species">Bdellovibrio bacteriovorus</named-content>

ABSTRACT Bdellovibrio bacteriovorus is a bacterial predator capable of killing and replicating inside most Gram-negative bacteria, including antibiotic-resistant pathogens. Despite growing interest in this organism as a potential therapeutic, many of its genes remain uncharacterized. Here, we perfor...

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Autores principales:	Miles C. Duncan, Rebecca K. Gillette, Micah A. Maglasang, Elizabeth A. Corn, Albert K. Tai, David W. Lazinski, Robert M. Q. Shanks, Daniel E. Kadouri, Andrew Camilli
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	Bdellovibrio bacteriovorus Escherichia coli Tn-seq Vibrio cholerae predatory bacteria Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/b8b6d2cbea3546999ddf7be7c9bfba0d
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spelling	oai:doaj.org-article:b8b6d2cbea3546999ddf7be7c9bfba0d2021-11-15T15:55:23ZHigh-Throughput Analysis of Gene Function in the Bacterial Predator <named-content content-type="genus-species">Bdellovibrio bacteriovorus</named-content>10.1128/mBio.01040-192150-7511https://doaj.org/article/b8b6d2cbea3546999ddf7be7c9bfba0d2019-06-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01040-19https://doaj.org/toc/2150-7511ABSTRACT Bdellovibrio bacteriovorus is a bacterial predator capable of killing and replicating inside most Gram-negative bacteria, including antibiotic-resistant pathogens. Despite growing interest in this organism as a potential therapeutic, many of its genes remain uncharacterized. Here, we perform a high-throughput genetic screen with B. bacteriovorus using transposon sequencing (Tn-seq) to explore the genetic requirements of predation. Two hundred one genes were deemed essential for growth in the absence of prey, whereas over 100 genes were found to be specifically required for predative growth on the human pathogens Vibrio cholerae and Escherichia coli in both planktonic and biofilm states. To further this work, we created an ordered-knockout library in B. bacteriovorus and developed new high-throughput techniques to characterize the mutants by their stage of deficiency in the predator life cycle. Using microscopy and flow cytometry, we confirmed 10 mutants defective in prey attachment and eight mutants defective in prey rounding. The majority of these genes are hypothetical and previously uncharacterized. Finally, we propose new nomenclature to group B. bacteriovorus mutants into classes based on their stage of predation defect. These results contribute to our basic understanding of bacterial predation and may be useful for harnessing B. bacteriovorus to kill harmful pathogens in the clinical setting. IMPORTANCE Bdellovibrio bacteriovorus is a predatory bacterium that can kill a wide range of Gram-negative bacteria, including many human pathogens. Given the global rise of antibiotic resistance and dearth of new antibiotics discovered in the past 30 years, this predator has potential as an alternative to traditional antibiotics. For many years, B. bacteriovorus research was hampered by a lack of genetic tools, and the genetic mechanisms of predation have only recently begun to be established. Here, we comprehensively identify and characterize predator genes required for killing bacterial prey, as well as genes that interfere in this process, which may allow us to design better therapeutic predators. Based on our study, we and other researchers may ultimately be able to genetically engineer strains that have improved killing rates, target specific species of prey, or preferentially target prey in the planktonic or biofilm state.Miles C. DuncanRebecca K. GilletteMicah A. MaglasangElizabeth A. CornAlbert K. TaiDavid W. LazinskiRobert M. Q. ShanksDaniel E. KadouriAndrew CamilliAmerican Society for MicrobiologyarticleBdellovibrio bacteriovorusEscherichia coliTn-seqVibrio choleraepredatory bacteriaMicrobiologyQR1-502ENmBio, Vol 10, Iss 3 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	Bdellovibrio bacteriovorus Escherichia coli Tn-seq Vibrio cholerae predatory bacteria Microbiology QR1-502
spellingShingle	Bdellovibrio bacteriovorus Escherichia coli Tn-seq Vibrio cholerae predatory bacteria Microbiology QR1-502 Miles C. Duncan Rebecca K. Gillette Micah A. Maglasang Elizabeth A. Corn Albert K. Tai David W. Lazinski Robert M. Q. Shanks Daniel E. Kadouri Andrew Camilli High-Throughput Analysis of Gene Function in the Bacterial Predator <named-content content-type="genus-species">Bdellovibrio bacteriovorus</named-content>
description	ABSTRACT Bdellovibrio bacteriovorus is a bacterial predator capable of killing and replicating inside most Gram-negative bacteria, including antibiotic-resistant pathogens. Despite growing interest in this organism as a potential therapeutic, many of its genes remain uncharacterized. Here, we perform a high-throughput genetic screen with B. bacteriovorus using transposon sequencing (Tn-seq) to explore the genetic requirements of predation. Two hundred one genes were deemed essential for growth in the absence of prey, whereas over 100 genes were found to be specifically required for predative growth on the human pathogens Vibrio cholerae and Escherichia coli in both planktonic and biofilm states. To further this work, we created an ordered-knockout library in B. bacteriovorus and developed new high-throughput techniques to characterize the mutants by their stage of deficiency in the predator life cycle. Using microscopy and flow cytometry, we confirmed 10 mutants defective in prey attachment and eight mutants defective in prey rounding. The majority of these genes are hypothetical and previously uncharacterized. Finally, we propose new nomenclature to group B. bacteriovorus mutants into classes based on their stage of predation defect. These results contribute to our basic understanding of bacterial predation and may be useful for harnessing B. bacteriovorus to kill harmful pathogens in the clinical setting. IMPORTANCE Bdellovibrio bacteriovorus is a predatory bacterium that can kill a wide range of Gram-negative bacteria, including many human pathogens. Given the global rise of antibiotic resistance and dearth of new antibiotics discovered in the past 30 years, this predator has potential as an alternative to traditional antibiotics. For many years, B. bacteriovorus research was hampered by a lack of genetic tools, and the genetic mechanisms of predation have only recently begun to be established. Here, we comprehensively identify and characterize predator genes required for killing bacterial prey, as well as genes that interfere in this process, which may allow us to design better therapeutic predators. Based on our study, we and other researchers may ultimately be able to genetically engineer strains that have improved killing rates, target specific species of prey, or preferentially target prey in the planktonic or biofilm state.
format	article
author	Miles C. Duncan Rebecca K. Gillette Micah A. Maglasang Elizabeth A. Corn Albert K. Tai David W. Lazinski Robert M. Q. Shanks Daniel E. Kadouri Andrew Camilli
author_facet	Miles C. Duncan Rebecca K. Gillette Micah A. Maglasang Elizabeth A. Corn Albert K. Tai David W. Lazinski Robert M. Q. Shanks Daniel E. Kadouri Andrew Camilli
author_sort	Miles C. Duncan
title	High-Throughput Analysis of Gene Function in the Bacterial Predator <named-content content-type="genus-species">Bdellovibrio bacteriovorus</named-content>
title_short	High-Throughput Analysis of Gene Function in the Bacterial Predator <named-content content-type="genus-species">Bdellovibrio bacteriovorus</named-content>
title_full	High-Throughput Analysis of Gene Function in the Bacterial Predator <named-content content-type="genus-species">Bdellovibrio bacteriovorus</named-content>
title_fullStr	High-Throughput Analysis of Gene Function in the Bacterial Predator <named-content content-type="genus-species">Bdellovibrio bacteriovorus</named-content>
title_full_unstemmed	High-Throughput Analysis of Gene Function in the Bacterial Predator <named-content content-type="genus-species">Bdellovibrio bacteriovorus</named-content>
title_sort	high-throughput analysis of gene function in the bacterial predator <named-content content-type="genus-species">bdellovibrio bacteriovorus</named-content>
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/b8b6d2cbea3546999ddf7be7c9bfba0d
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High-Throughput Analysis of Gene Function in the Bacterial Predator <named-content content-type="genus-species">Bdellovibrio bacteriovorus</named-content>

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