Complex Multilevel Control of Hemolysin Production by Uropathogenic <named-content content-type="genus-species">Escherichia coli</named-content>

ABSTRACT Uropathogenic Escherichia coli (UPEC) is the major cause of urinary tract infections. Nearly half of all UPEC strains secrete hemolysin, a cytotoxic pore-forming toxin. Here, we show that the prevalence of the hemolysin toxin gene (hlyA) is highly variable among the most common 83 E. coli s...

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Autores principales:	Nguyen Thi Khanh Nhu, Minh-Duy Phan, Brian M. Forde, Ambika M. V. Murthy, Kate M. Peters, Christopher J. Day, Jessica Poole, Timothy J. Kidd, Rodney A. Welch, Michael P. Jennings, Glen C. Ulett, Matthew J. Sweet, Scott A. Beatson, Mark A. Schembri
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	Escherichia coli TraDIS gene regulation hemolysin urinary tract infection virulence Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/5d5d70f495de450ea3ff96ceda70c3ec
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spelling	oai:doaj.org-article:5d5d70f495de450ea3ff96ceda70c3ec2021-11-15T15:59:41ZComplex Multilevel Control of Hemolysin Production by Uropathogenic <named-content content-type="genus-species">Escherichia coli</named-content>10.1128/mBio.02248-192150-7511https://doaj.org/article/5d5d70f495de450ea3ff96ceda70c3ec2019-10-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.02248-19https://doaj.org/toc/2150-7511ABSTRACT Uropathogenic Escherichia coli (UPEC) is the major cause of urinary tract infections. Nearly half of all UPEC strains secrete hemolysin, a cytotoxic pore-forming toxin. Here, we show that the prevalence of the hemolysin toxin gene (hlyA) is highly variable among the most common 83 E. coli sequence types (STs) represented on the EnteroBase genome database. To explore this diversity in the context of a defined monophyletic lineage, we contextualized sequence variation of the hlyCABD operon within the genealogy of the globally disseminated multidrug-resistant ST131 clone. We show that sequence changes in hlyCABD and its newly defined 1.616-kb-long leader sequence correspond to phylogenetic designation, and that ST131 strains with the strongest hemolytic activity belong to the most extensive multidrug-resistant sublineage (clade C2). To define the set of genes involved in hemolysin production, the clade C2 strain S65EC was completely sequenced and subjected to a genome-wide screen by combining saturated transposon mutagenesis and transposon-directed insertion site sequencing with the capacity to lyse red blood cells. Using this approach, and subsequent targeted mutagenesis and complementation, 13 genes were confirmed to be specifically required for production of active hemolysin. New hemolysin-controlling elements included discrete sets of genes involved in lipopolysaccharide (LPS) inner core biosynthesis (waaC, waaF, waaG, and rfaE) and cytoplasmic chaperone activity (dnaK and dnaJ), and we show these are required for hemolysin secretion. Overall, this work provides a unique description of hemolysin sequence diversity in a single clonal lineage and describes a complex multilevel system of regulatory control for this important toxin. IMPORTANCE Uropathogenic E. coli (UPEC) is the major cause of urinary tract infections and a frequent cause of sepsis. Nearly half of all UPEC strains produce the potent cytotoxin hemolysin, and its expression is associated with enhanced virulence. In this study, we explored hemolysin variation within the globally dominant UPEC ST131 clone, finding that strains from the ST131 sublineage with the greatest multidrug resistance also possess the strongest hemolytic activity. We also employed an innovative forward genetic screen to define the set of genes required for hemolysin production. Using this approach, and subsequent targeted mutagenesis and complementation, we identified new hemolysin-controlling elements involved in LPS inner core biosynthesis and cytoplasmic chaperone activity, and we show that mechanistically they are required for hemolysin secretion. These original discoveries substantially enhance our understanding of hemolysin regulation, secretion and function.Nguyen Thi Khanh NhuMinh-Duy PhanBrian M. FordeAmbika M. V. MurthyKate M. PetersChristopher J. DayJessica PooleTimothy J. KiddRodney A. WelchMichael P. JenningsGlen C. UlettMatthew J. SweetScott A. BeatsonMark A. SchembriAmerican Society for MicrobiologyarticleEscherichia coliTraDISgene regulationhemolysinurinary tract infectionvirulenceMicrobiologyQR1-502ENmBio, Vol 10, Iss 5 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	Escherichia coli TraDIS gene regulation hemolysin urinary tract infection virulence Microbiology QR1-502
spellingShingle	Escherichia coli TraDIS gene regulation hemolysin urinary tract infection virulence Microbiology QR1-502 Nguyen Thi Khanh Nhu Minh-Duy Phan Brian M. Forde Ambika M. V. Murthy Kate M. Peters Christopher J. Day Jessica Poole Timothy J. Kidd Rodney A. Welch Michael P. Jennings Glen C. Ulett Matthew J. Sweet Scott A. Beatson Mark A. Schembri Complex Multilevel Control of Hemolysin Production by Uropathogenic <named-content content-type="genus-species">Escherichia coli</named-content>
description	ABSTRACT Uropathogenic Escherichia coli (UPEC) is the major cause of urinary tract infections. Nearly half of all UPEC strains secrete hemolysin, a cytotoxic pore-forming toxin. Here, we show that the prevalence of the hemolysin toxin gene (hlyA) is highly variable among the most common 83 E. coli sequence types (STs) represented on the EnteroBase genome database. To explore this diversity in the context of a defined monophyletic lineage, we contextualized sequence variation of the hlyCABD operon within the genealogy of the globally disseminated multidrug-resistant ST131 clone. We show that sequence changes in hlyCABD and its newly defined 1.616-kb-long leader sequence correspond to phylogenetic designation, and that ST131 strains with the strongest hemolytic activity belong to the most extensive multidrug-resistant sublineage (clade C2). To define the set of genes involved in hemolysin production, the clade C2 strain S65EC was completely sequenced and subjected to a genome-wide screen by combining saturated transposon mutagenesis and transposon-directed insertion site sequencing with the capacity to lyse red blood cells. Using this approach, and subsequent targeted mutagenesis and complementation, 13 genes were confirmed to be specifically required for production of active hemolysin. New hemolysin-controlling elements included discrete sets of genes involved in lipopolysaccharide (LPS) inner core biosynthesis (waaC, waaF, waaG, and rfaE) and cytoplasmic chaperone activity (dnaK and dnaJ), and we show these are required for hemolysin secretion. Overall, this work provides a unique description of hemolysin sequence diversity in a single clonal lineage and describes a complex multilevel system of regulatory control for this important toxin. IMPORTANCE Uropathogenic E. coli (UPEC) is the major cause of urinary tract infections and a frequent cause of sepsis. Nearly half of all UPEC strains produce the potent cytotoxin hemolysin, and its expression is associated with enhanced virulence. In this study, we explored hemolysin variation within the globally dominant UPEC ST131 clone, finding that strains from the ST131 sublineage with the greatest multidrug resistance also possess the strongest hemolytic activity. We also employed an innovative forward genetic screen to define the set of genes required for hemolysin production. Using this approach, and subsequent targeted mutagenesis and complementation, we identified new hemolysin-controlling elements involved in LPS inner core biosynthesis and cytoplasmic chaperone activity, and we show that mechanistically they are required for hemolysin secretion. These original discoveries substantially enhance our understanding of hemolysin regulation, secretion and function.
format	article
author	Nguyen Thi Khanh Nhu Minh-Duy Phan Brian M. Forde Ambika M. V. Murthy Kate M. Peters Christopher J. Day Jessica Poole Timothy J. Kidd Rodney A. Welch Michael P. Jennings Glen C. Ulett Matthew J. Sweet Scott A. Beatson Mark A. Schembri
author_facet	Nguyen Thi Khanh Nhu Minh-Duy Phan Brian M. Forde Ambika M. V. Murthy Kate M. Peters Christopher J. Day Jessica Poole Timothy J. Kidd Rodney A. Welch Michael P. Jennings Glen C. Ulett Matthew J. Sweet Scott A. Beatson Mark A. Schembri
author_sort	Nguyen Thi Khanh Nhu
title	Complex Multilevel Control of Hemolysin Production by Uropathogenic <named-content content-type="genus-species">Escherichia coli</named-content>
title_short	Complex Multilevel Control of Hemolysin Production by Uropathogenic <named-content content-type="genus-species">Escherichia coli</named-content>
title_full	Complex Multilevel Control of Hemolysin Production by Uropathogenic <named-content content-type="genus-species">Escherichia coli</named-content>
title_fullStr	Complex Multilevel Control of Hemolysin Production by Uropathogenic <named-content content-type="genus-species">Escherichia coli</named-content>
title_full_unstemmed	Complex Multilevel Control of Hemolysin Production by Uropathogenic <named-content content-type="genus-species">Escherichia coli</named-content>
title_sort	complex multilevel control of hemolysin production by uropathogenic <named-content content-type="genus-species">escherichia coli</named-content>
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/5d5d70f495de450ea3ff96ceda70c3ec
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Complex Multilevel Control of Hemolysin Production by Uropathogenic <named-content content-type="genus-species">Escherichia coli</named-content>

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