Staff View: Discovery of New Genes Involved in Curli Production by a Uropathogenic Escherichia coli Strain from the Highly Virulent O45:K1:H7 Lineage

Discovery of New Genes Involved in Curli Production by a Uropathogenic Escherichia coli Strain from the Highly Virulent O45:K1:H7 Lineage

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 show...

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Main Authors:	Nhu, Nguyen Thi Khanh, Phan, Minh-Duy, Peters, Kate M., Lo, Alvin W., Forde, Brian M., Chong, Teik Min, Yin, Wai Fong, Chan, Kok Gan, Chromek, Milan, Brauner, Annelie, Chapman, Matthew R., Beatson, Scott A., Schembri, Mark A., Justice, Sheryl
Format:	Article
Published:	American Society for Microbiology 2018
Subjects:	Q Science (General) QH Natural history
Online Access:	http://eprints.um.edu.my/21130/ https://doi.org/10.1128/mBio.01462-18
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spelling	my.um.eprints.211302019-05-07T03:35:42Z http://eprints.um.edu.my/21130/ Discovery of New Genes Involved in Curli Production by a Uropathogenic Escherichia coli Strain from the Highly Virulent O45:K1:H7 Lineage Nhu, Nguyen Thi Khanh Phan, Minh-Duy Peters, Kate M. Lo, Alvin W. Forde, Brian M. Chong, Teik Min Yin, Wai Fong Chan, Kok Gan Chromek, Milan Brauner, Annelie Chapman, Matthew R. Beatson, Scott A. Schembri, Mark A. Justice, Sheryl Q Science (General) QH Natural history 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 autotrans-porters. 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. American Society for Microbiology 2018 Article PeerReviewed Nhu, Nguyen Thi Khanh and Phan, Minh-Duy and Peters, Kate M. and Lo, Alvin W. and Forde, Brian M. and Chong, Teik Min and Yin, Wai Fong and Chan, Kok Gan and Chromek, Milan and Brauner, Annelie and Chapman, Matthew R. and Beatson, Scott A. and Schembri, Mark A. and Justice, Sheryl (2018) Discovery of New Genes Involved in Curli Production by a Uropathogenic Escherichia coli Strain from the Highly Virulent O45:K1:H7 Lineage. mBio, 9 (4). e01462-18. ISSN 2150-7511 https://doi.org/10.1128/mBio.01462-18 doi:10.1128/mBio.01462-18
institution	Universiti Malaya
building	UM Library
collection	Institutional Repository
continent	Asia
country	Malaysia
content_provider	Universiti Malaya
content_source	UM Research Repository
url_provider	http://eprints.um.edu.my/
topic	Q Science (General) QH Natural history
spellingShingle	Q Science (General) QH Natural history Nhu, Nguyen Thi Khanh Phan, Minh-Duy Peters, Kate M. Lo, Alvin W. Forde, Brian M. Chong, Teik Min Yin, Wai Fong Chan, Kok Gan Chromek, Milan Brauner, Annelie Chapman, Matthew R. Beatson, Scott A. Schembri, Mark A. Justice, Sheryl Discovery of New Genes Involved in Curli Production by a Uropathogenic Escherichia coli Strain from the Highly Virulent O45:K1:H7 Lineage
description	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 autotrans-porters. 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	Nhu, Nguyen Thi Khanh Phan, Minh-Duy Peters, Kate M. Lo, Alvin W. Forde, Brian M. Chong, Teik Min Yin, Wai Fong Chan, Kok Gan Chromek, Milan Brauner, Annelie Chapman, Matthew R. Beatson, Scott A. Schembri, Mark A. Justice, Sheryl
author_facet	Nhu, Nguyen Thi Khanh Phan, Minh-Duy Peters, Kate M. Lo, Alvin W. Forde, Brian M. Chong, Teik Min Yin, Wai Fong Chan, Kok Gan Chromek, Milan Brauner, Annelie Chapman, Matthew R. Beatson, Scott A. Schembri, Mark A. Justice, Sheryl
author_sort	Nhu, Nguyen Thi Khanh
title	Discovery of New Genes Involved in Curli Production by a Uropathogenic Escherichia coli Strain from the Highly Virulent O45:K1:H7 Lineage
title_short	Discovery of New Genes Involved in Curli Production by a Uropathogenic Escherichia coli Strain from the Highly Virulent O45:K1:H7 Lineage
title_full	Discovery of New Genes Involved in Curli Production by a Uropathogenic Escherichia coli Strain from the Highly Virulent O45:K1:H7 Lineage
title_fullStr	Discovery of New Genes Involved in Curli Production by a Uropathogenic Escherichia coli Strain from the Highly Virulent O45:K1:H7 Lineage
title_full_unstemmed	Discovery of New Genes Involved in Curli Production by a Uropathogenic Escherichia coli Strain from the Highly Virulent O45:K1:H7 Lineage
title_sort	discovery of new genes involved in curli production by a uropathogenic escherichia coli strain from the highly virulent o45:k1:h7 lineage
publisher	American Society for Microbiology
publishDate	2018
url	http://eprints.um.edu.my/21130/ https://doi.org/10.1128/mBio.01462-18
_version_	1643691474612649984
score	13.211869

Discovery of New Genes Involved in Curli Production by a Uropathogenic Escherichia coli Strain from the Highly Virulent O45:K1:H7 Lineage

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