Discovering the Molecular Determinants of <named-content content-type="genus-species">Phaeobacter inhibens</named-content> Susceptibility to Phaeobacter Phage MD18

ABSTRACT Bacteriophages have immense potential as antibiotic therapies and in genetic engineering. Understanding the mechanisms that bacteriophages implement to infect their hosts will allow researchers to manipulate these systems and adapt them to specific bacterial targets. In this study, we isola...

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Autores principales:	Guillaume Urtecho, Danielle E. Campbell, David M. Hershey, Fatima A. Hussain, Rachel J. Whitaker, George A. O’Toole
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	bacteriophage BarSeq ecology microbial interactions phage genomics Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/9eee29da79544916bea4fc57ce355514
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spelling	oai:doaj.org-article:9eee29da79544916bea4fc57ce3555142021-11-15T15:31:13ZDiscovering the Molecular Determinants of <named-content content-type="genus-species">Phaeobacter inhibens</named-content> Susceptibility to Phaeobacter Phage MD1810.1128/mSphere.00898-202379-5042https://doaj.org/article/9eee29da79544916bea4fc57ce3555142020-12-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00898-20https://doaj.org/toc/2379-5042ABSTRACT Bacteriophages have immense potential as antibiotic therapies and in genetic engineering. Understanding the mechanisms that bacteriophages implement to infect their hosts will allow researchers to manipulate these systems and adapt them to specific bacterial targets. In this study, we isolated a bacteriophage capable of infecting the marine alphaproteobacterium Phaeobacter inhibens and determined its mechanism of infection. Phaeobacter virus MD18, a novel species of bacteriophage isolated in Woods Hole, MA, exhibits potent lytic ability against P. inhibens and appears to be of the Siphoviridae morphotype. The genomic sequence of MD18 displayed significant similarity to another siphophage, the recently discovered Roseobacter phage DSS3P8, but genomic and phylogenetic analyses, assessing host range and a search of available metagenomes are all consistent with the conclusion that Phaeobacter phage MD18 is a novel lytic phage. We incubated MD18 with a library of barcoded P. inhibens transposon insertion mutants and identified 22 genes that appear to be required for phage predation of this host. Network analysis of these genes using genomic position, Gene Ontology (GO) term enrichment, and protein associations revealed that these genes are enriched for roles in assembly of a type IV pilus (T4P) and regulators of cellular morphology. Our results suggest that T4P serve as receptors for a novel marine virus that targets P. inhibens. IMPORTANCE Bacteriophages are useful nonantibiotic therapeutics for bacterial infections as well as threats to industries utilizing bacterial agents. This study identified Phaeobacter virus MD18, a phage antagonist of Phaeobacter inhibens, a bacterium with promising use as a probiotic for aquatic farming industries. Genomic analysis suggested that Phaeobacter phage MD18 has evolved to enhance its replication in P. inhibens by adopting favorable tRNA genes as well as through genomic sequence adaptation to resemble host codon usage. Lastly, a high-throughput analysis of P. inhibens transposon insertion mutants identified genes that modulate host susceptibility to phage MD18 and implicated the type IV pilus as the likely receptor recognized for adsorption. This study marks the first characterization of the relationship between P. inhibens and an environmentally sampled phage, which informs our understanding of natural threats to the bacterium and may promote the development of novel phage technologies for genetic manipulation of this host.Guillaume UrtechoDanielle E. CampbellDavid M. HersheyFatima A. HussainRachel J. WhitakerGeorge A. O’TooleAmerican Society for MicrobiologyarticlebacteriophageBarSeqecologymicrobial interactionsphage genomicsMicrobiologyQR1-502ENmSphere, Vol 5, Iss 6 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	bacteriophage BarSeq ecology microbial interactions phage genomics Microbiology QR1-502
spellingShingle	bacteriophage BarSeq ecology microbial interactions phage genomics Microbiology QR1-502 Guillaume Urtecho Danielle E. Campbell David M. Hershey Fatima A. Hussain Rachel J. Whitaker George A. O’Toole Discovering the Molecular Determinants of <named-content content-type="genus-species">Phaeobacter inhibens</named-content> Susceptibility to Phaeobacter Phage MD18
description	ABSTRACT Bacteriophages have immense potential as antibiotic therapies and in genetic engineering. Understanding the mechanisms that bacteriophages implement to infect their hosts will allow researchers to manipulate these systems and adapt them to specific bacterial targets. In this study, we isolated a bacteriophage capable of infecting the marine alphaproteobacterium Phaeobacter inhibens and determined its mechanism of infection. Phaeobacter virus MD18, a novel species of bacteriophage isolated in Woods Hole, MA, exhibits potent lytic ability against P. inhibens and appears to be of the Siphoviridae morphotype. The genomic sequence of MD18 displayed significant similarity to another siphophage, the recently discovered Roseobacter phage DSS3P8, but genomic and phylogenetic analyses, assessing host range and a search of available metagenomes are all consistent with the conclusion that Phaeobacter phage MD18 is a novel lytic phage. We incubated MD18 with a library of barcoded P. inhibens transposon insertion mutants and identified 22 genes that appear to be required for phage predation of this host. Network analysis of these genes using genomic position, Gene Ontology (GO) term enrichment, and protein associations revealed that these genes are enriched for roles in assembly of a type IV pilus (T4P) and regulators of cellular morphology. Our results suggest that T4P serve as receptors for a novel marine virus that targets P. inhibens. IMPORTANCE Bacteriophages are useful nonantibiotic therapeutics for bacterial infections as well as threats to industries utilizing bacterial agents. This study identified Phaeobacter virus MD18, a phage antagonist of Phaeobacter inhibens, a bacterium with promising use as a probiotic for aquatic farming industries. Genomic analysis suggested that Phaeobacter phage MD18 has evolved to enhance its replication in P. inhibens by adopting favorable tRNA genes as well as through genomic sequence adaptation to resemble host codon usage. Lastly, a high-throughput analysis of P. inhibens transposon insertion mutants identified genes that modulate host susceptibility to phage MD18 and implicated the type IV pilus as the likely receptor recognized for adsorption. This study marks the first characterization of the relationship between P. inhibens and an environmentally sampled phage, which informs our understanding of natural threats to the bacterium and may promote the development of novel phage technologies for genetic manipulation of this host.
format	article
author	Guillaume Urtecho Danielle E. Campbell David M. Hershey Fatima A. Hussain Rachel J. Whitaker George A. O’Toole
author_facet	Guillaume Urtecho Danielle E. Campbell David M. Hershey Fatima A. Hussain Rachel J. Whitaker George A. O’Toole
author_sort	Guillaume Urtecho
title	Discovering the Molecular Determinants of <named-content content-type="genus-species">Phaeobacter inhibens</named-content> Susceptibility to Phaeobacter Phage MD18
title_short	Discovering the Molecular Determinants of <named-content content-type="genus-species">Phaeobacter inhibens</named-content> Susceptibility to Phaeobacter Phage MD18
title_full	Discovering the Molecular Determinants of <named-content content-type="genus-species">Phaeobacter inhibens</named-content> Susceptibility to Phaeobacter Phage MD18
title_fullStr	Discovering the Molecular Determinants of <named-content content-type="genus-species">Phaeobacter inhibens</named-content> Susceptibility to Phaeobacter Phage MD18
title_full_unstemmed	Discovering the Molecular Determinants of <named-content content-type="genus-species">Phaeobacter inhibens</named-content> Susceptibility to Phaeobacter Phage MD18
title_sort	discovering the molecular determinants of <named-content content-type="genus-species">phaeobacter inhibens</named-content> susceptibility to phaeobacter phage md18
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/9eee29da79544916bea4fc57ce355514
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Discovering the Molecular Determinants of <named-content content-type="genus-species">Phaeobacter inhibens</named-content> Susceptibility to Phaeobacter Phage MD18

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