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dc.contributor.authorNhu, Nguyen Thi Khanh
dc.contributor.authorPhan, Minh-Duy
dc.contributor.authorForde, Brian M
dc.contributor.authorMurthy, Ambika MV
dc.contributor.authorPeters, Kate M
dc.contributor.authorDay, Christopher J
dc.contributor.authorPoole, Jessica
dc.contributor.authorKidd, Timothy J
dc.contributor.authorWelch, Rodney A
dc.contributor.authorJennings, Michael P
dc.contributor.authorUlett, Glen C
dc.contributor.authorSweet, Matthew J
dc.contributor.authorBeatson, Scott A
dc.contributor.authorSchembri, Mark A
dc.date.accessioned2019-10-09T23:52:08Z
dc.date.available2019-10-09T23:52:08Z
dc.date.issued2019
dc.identifier.issn2161-2129
dc.identifier.doi10.1128/mBio.02248-19
dc.identifier.urihttp://hdl.handle.net/10072/388206
dc.description.abstractUropathogenic 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.
dc.description.peerreviewedYes
dc.languageEnglish
dc.publisherAmerican Society for Microbiology
dc.publisher.placeUnited States
dc.relation.ispartofpagefrome02248: 1
dc.relation.ispartofpagetoe02248: 17
dc.relation.ispartofissue5
dc.relation.ispartofjournalMBio
dc.relation.ispartofvolume10
dc.subject.fieldofresearchMicrobiology
dc.subject.fieldofresearchcode0605
dc.subject.keywordsEscherichia coli
dc.subject.keywordsTraDIS
dc.subject.keywordsgene regulation
dc.subject.keywordshemolysin
dc.subject.keywordsurinary tract infection
dc.titleComplex Multilevel Control of Hemolysin Production by Uropathogenic Escherichia coli
dc.typeJournal article
dc.type.descriptionC1 - Articles
dcterms.bibliographicCitationNhu, NTK; Phan, M-D; Forde, BM; Murthy, AMV; Peters, KM; Day, CJ; Poole, J; Kidd, TJ; Welch, RA; Jennings, MP; Ulett, GC; Sweet, MJ; Beatson, SA; Schembri, MA, Complex Multilevel Control of Hemolysin Production by Uropathogenic Escherichia coli., MBio, 2019, 10 (5), pp. e02248: 1-e02248: 17
dcterms.licensehttps://creativecommons.org/licenses/by/4.0/
dc.date.updated2019-10-09T23:33:18Z
dc.description.versionPublished
gro.rights.copyrightCopyright © 2019 Nhu et al. This is an Open Access article distributed under the terms of the Creative Commons Attribution 4.0 International License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
gro.hasfulltextFull Text
gro.griffith.authorUlett, Glen C.
gro.griffith.authorDay, Christopher J.
gro.griffith.authorPoole, Jessica
gro.griffith.authorJennings, Michael P.


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