Discovery of New Genes Involved in Curli Production by a Uropathogenic <named-content content-type="genus-species">Escherichia coli</named-content> Strain from the Highly Virulent O45:K1:H7 Lineage

ABSTRACT Curli are bacterial surface-associated amyloid fibers that bind to the dye Congo red (CR) and facilitate uropathogenic Escherichia coli (UPEC) biofilm formation and protection against host innate defenses. Here we sequenced the genome of the curli-producing UPEC pyelonephritis strain MS7163...

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Autores principales:	Nguyen Thi Khanh Nhu, Minh-Duy Phan, Kate M. Peters, Alvin W. Lo, Brian M. Forde, Teik Min Chong, Wai-Fong Yin, Kok-Gan Chan, Milan Chromek, Annelie Brauner, Matthew R. Chapman, Scott A. Beatson, Mark A. Schembri
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2018
Materias:	Escherichia coli biofilms curli urinary tract infection virulence virulence regulation Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/a037aae2567348088f8d6a518a6ad468
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spelling	oai:doaj.org-article:a037aae2567348088f8d6a518a6ad4682021-11-15T16:00:14ZDiscovery of New Genes Involved in Curli Production by a Uropathogenic <named-content content-type="genus-species">Escherichia coli</named-content> Strain from the Highly Virulent O45:K1:H7 Lineage10.1128/mBio.01462-182150-7511https://doaj.org/article/a037aae2567348088f8d6a518a6ad4682018-09-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01462-18https://doaj.org/toc/2150-7511ABSTRACT Curli are bacterial surface-associated amyloid fibers that bind to the dye Congo red (CR) and facilitate uropathogenic Escherichia coli (UPEC) biofilm formation and protection against host innate defenses. Here we sequenced the genome of the curli-producing UPEC pyelonephritis strain MS7163 and showed it belongs to the highly virulent O45:K1:H7 neonatal meningitis-associated clone. MS7163 produced curli at human physiological temperature, and this correlated with biofilm growth, resistance of sessile cells to the human cationic peptide cathelicidin, and enhanced colonization of the mouse bladder. We devised a forward genetic screen using CR staining as a proxy for curli production and identified 41 genes that were required for optimal CR binding, of which 19 genes were essential for curli synthesis. Ten of these genes were novel or poorly characterized with respect to curli synthesis and included genes involved in purine de novo biosynthesis, a regulator that controls the Rcs phosphorelay system, and a novel repressor of curli production (referred to as rcpA). The involvement of these genes in curli production was confirmed by the construction of defined mutants and their complementation. The mutants did not express the curli major subunit CsgA and failed to produce curli based on CR binding. Mutation of purF (the first gene in the purine biosynthesis pathway) and rcpA also led to attenuated colonization of the mouse bladder. Overall, this work has provided new insight into the regulation of curli and the role of these amyloid fibers in UPEC biofilm formation and pathogenesis. IMPORTANCE Uropathogenic Escherichia coli (UPEC) strains are the most common cause of urinary tract infection, a disease increasingly associated with escalating antibiotic resistance. UPEC strains possess multiple surface-associated factors that enable their colonization of the urinary tract, including fimbriae, curli, and autotransporters. Curli are extracellular amyloid fibers that enhance UPEC virulence and promote biofilm formation. Here we examined the function and regulation of curli in a UPEC pyelonephritis strain belonging to the highly virulent O45:K1:H7 neonatal meningitis-associated clone. Curli expression at human physiological temperature led to increased biofilm formation, resistance of sessile cells to the human cationic peptide LL-37, and enhanced bladder colonization. Using a comprehensive genetic screen, we identified multiple genes involved in curli production, including several that were novel or poorly characterized with respect to curli synthesis. In total, this study demonstrates an important role for curli as a UPEC virulence factor that promotes biofilm formation, resistance, and pathogenesis.Nguyen Thi Khanh NhuMinh-Duy PhanKate M. PetersAlvin W. LoBrian M. FordeTeik Min ChongWai-Fong YinKok-Gan ChanMilan ChromekAnnelie BraunerMatthew R. ChapmanScott A. BeatsonMark A. SchembriAmerican Society for MicrobiologyarticleEscherichia colibiofilmscurliurinary tract infectionvirulencevirulence regulationMicrobiologyQR1-502ENmBio, Vol 9, Iss 4 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Escherichia coli biofilms curli urinary tract infection virulence virulence regulation Microbiology QR1-502
spellingShingle	Escherichia coli biofilms curli urinary tract infection virulence virulence regulation Microbiology QR1-502 Nguyen Thi Khanh Nhu Minh-Duy Phan Kate M. Peters Alvin W. Lo Brian M. Forde Teik Min Chong Wai-Fong Yin Kok-Gan Chan Milan Chromek Annelie Brauner Matthew R. Chapman Scott A. Beatson Mark A. Schembri Discovery of New Genes Involved in Curli Production by a Uropathogenic <named-content content-type="genus-species">Escherichia coli</named-content> Strain from the Highly Virulent O45:K1:H7 Lineage
description	ABSTRACT Curli are bacterial surface-associated amyloid fibers that bind to the dye Congo red (CR) and facilitate uropathogenic Escherichia coli (UPEC) biofilm formation and protection against host innate defenses. Here we sequenced the genome of the curli-producing UPEC pyelonephritis strain MS7163 and showed it belongs to the highly virulent O45:K1:H7 neonatal meningitis-associated clone. MS7163 produced curli at human physiological temperature, and this correlated with biofilm growth, resistance of sessile cells to the human cationic peptide cathelicidin, and enhanced colonization of the mouse bladder. We devised a forward genetic screen using CR staining as a proxy for curli production and identified 41 genes that were required for optimal CR binding, of which 19 genes were essential for curli synthesis. Ten of these genes were novel or poorly characterized with respect to curli synthesis and included genes involved in purine de novo biosynthesis, a regulator that controls the Rcs phosphorelay system, and a novel repressor of curli production (referred to as rcpA). The involvement of these genes in curli production was confirmed by the construction of defined mutants and their complementation. The mutants did not express the curli major subunit CsgA and failed to produce curli based on CR binding. Mutation of purF (the first gene in the purine biosynthesis pathway) and rcpA also led to attenuated colonization of the mouse bladder. Overall, this work has provided new insight into the regulation of curli and the role of these amyloid fibers in UPEC biofilm formation and pathogenesis. IMPORTANCE Uropathogenic Escherichia coli (UPEC) strains are the most common cause of urinary tract infection, a disease increasingly associated with escalating antibiotic resistance. UPEC strains possess multiple surface-associated factors that enable their colonization of the urinary tract, including fimbriae, curli, and autotransporters. Curli are extracellular amyloid fibers that enhance UPEC virulence and promote biofilm formation. Here we examined the function and regulation of curli in a UPEC pyelonephritis strain belonging to the highly virulent O45:K1:H7 neonatal meningitis-associated clone. Curli expression at human physiological temperature led to increased biofilm formation, resistance of sessile cells to the human cationic peptide LL-37, and enhanced bladder colonization. Using a comprehensive genetic screen, we identified multiple genes involved in curli production, including several that were novel or poorly characterized with respect to curli synthesis. In total, this study demonstrates an important role for curli as a UPEC virulence factor that promotes biofilm formation, resistance, and pathogenesis.
format	article
author	Nguyen Thi Khanh Nhu Minh-Duy Phan Kate M. Peters Alvin W. Lo Brian M. Forde Teik Min Chong Wai-Fong Yin Kok-Gan Chan Milan Chromek Annelie Brauner Matthew R. Chapman Scott A. Beatson Mark A. Schembri
author_facet	Nguyen Thi Khanh Nhu Minh-Duy Phan Kate M. Peters Alvin W. Lo Brian M. Forde Teik Min Chong Wai-Fong Yin Kok-Gan Chan Milan Chromek Annelie Brauner Matthew R. Chapman Scott A. Beatson Mark A. Schembri
author_sort	Nguyen Thi Khanh Nhu
title	Discovery of New Genes Involved in Curli Production by a Uropathogenic <named-content content-type="genus-species">Escherichia coli</named-content> Strain from the Highly Virulent O45:K1:H7 Lineage
title_short	Discovery of New Genes Involved in Curli Production by a Uropathogenic <named-content content-type="genus-species">Escherichia coli</named-content> Strain from the Highly Virulent O45:K1:H7 Lineage
title_full	Discovery of New Genes Involved in Curli Production by a Uropathogenic <named-content content-type="genus-species">Escherichia coli</named-content> Strain from the Highly Virulent O45:K1:H7 Lineage
title_fullStr	Discovery of New Genes Involved in Curli Production by a Uropathogenic <named-content content-type="genus-species">Escherichia coli</named-content> Strain from the Highly Virulent O45:K1:H7 Lineage
title_full_unstemmed	Discovery of New Genes Involved in Curli Production by a Uropathogenic <named-content content-type="genus-species">Escherichia coli</named-content> Strain from the Highly Virulent O45:K1:H7 Lineage
title_sort	discovery of new genes involved in curli production by a uropathogenic <named-content content-type="genus-species">escherichia coli</named-content> strain from the highly virulent o45:k1:h7 lineage
publisher	American Society for Microbiology
publishDate	2018
url	https://doaj.org/article/a037aae2567348088f8d6a518a6ad468
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Discovery of New Genes Involved in Curli Production by a Uropathogenic <named-content content-type="genus-species">Escherichia coli</named-content> Strain from the Highly Virulent O45:K1:H7 Lineage

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