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dc.contributor.authorGoh, Kelvin GK
dc.contributor.authorMinh-Duy, Phan
dc.contributor.authorForde, Brian M
dc.contributor.authorChong, Teik Min
dc.contributor.authorYin, Wai-Fong
dc.contributor.authorChan, Kok-Gan
dc.contributor.authorUlett, Glen C
dc.contributor.authorSweet, Matthew J
dc.contributor.authorBeatson, Scott A
dc.contributor.authorSchembri, Mark A
dc.date.accessioned2018-03-08T04:07:46Z
dc.date.available2018-03-08T04:07:46Z
dc.date.issued2017
dc.identifier.issn2150-7511
dc.identifier.doi10.1128/mBio.01558-17
dc.identifier.urihttp://hdl.handle.net/10072/370781
dc.description.abstractUropathogenic Escherichia coli (UPEC) is a major cause of urinary tract and bloodstream infections and possesses an array of virulence factors for colonization, survival, and persistence. One such factor is the polysaccharide K capsule. Among the different K capsule types, the K1 serotype is strongly associated with UPEC infection. In this study, we completely sequenced the K1 UPEC urosepsis strain PA45B and employed a novel combination of a lytic K1 capsule-specific phage, saturated Tn5 transposon mutagenesis, and high-throughput transposon-directed insertion site sequencing (TraDIS) to identify the complement of genes required for capsule production. Our analysis identified known genes involved in capsule biosynthesis, as well as two additional regulatory genes (mprA and lrhA) that we characterized at the molecular level. Mutation of mprA resulted in protection against K1 phage-mediated killing, a phenotype restored by complementation. We also identified a significantly increased unidirectional Tn5 insertion frequency upstream of the lrhA gene and showed that strong expression of LrhA induced by a constitutive Pcl promoter led to loss of capsule production. Further analysis revealed loss of MprA or overexpression of LrhA affected the transcription of capsule biosynthesis genes in PA45B and increased sensitivity to killing in whole blood. Similar phenotypes were also observed in UPEC strains UTI89 (K1) and CFT073 (K2), demonstrating that the effects were neither strain nor capsule type specific. Overall, this study defined the genome of a UPEC urosepsis isolate and identified and characterized two new regulatory factors that affect UPEC capsule production.
dc.description.peerreviewedYes
dc.languageEnglish
dc.language.isoeng
dc.publisherAmerican Society for Microbiology
dc.relation.ispartofpagefrome01558-17-1
dc.relation.ispartofpagetoe01558-17-16
dc.relation.ispartofissue5
dc.relation.ispartofjournalmBio
dc.relation.ispartofvolume8
dc.subject.fieldofresearchMicrobiology
dc.subject.fieldofresearchMicrobiology not elsewhere classified
dc.subject.fieldofresearchBiochemistry and cell biology
dc.subject.fieldofresearchMedical microbiology
dc.subject.fieldofresearchcode3107
dc.subject.fieldofresearchcode310799
dc.subject.fieldofresearchcode3101
dc.subject.fieldofresearchcode3207
dc.titleGenome-Wide Discovery of Genes Required for Capsule Production by Uropathogenic Escherichia coli
dc.typeJournal article
dc.type.descriptionC1 - Articles
dc.type.codeC - Journal Articles
dcterms.licensehttp://creativecommons.org/licenses/by/4.0/
dc.description.versionVersion of Record (VoR)
gro.rights.copyright© 2017 Goh et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (http://creativecommons.org/licenses/by/4.0/) 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.


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