scholarly journals Identification and Characterization of a Phase-Variable Element That Regulates the Autotransporter UpaE in UropathogenicEscherichia coli

mBio ◽  
2018 ◽  
Vol 9 (4) ◽  
Author(s):  
E. J. Battaglioli ◽  
K. G. K. Goh ◽  
T. S. Atruktsang ◽  
K. Schwartz ◽  
M. A. Schembri ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Extracellular Matrix ◽  
Urinary Tract Infection ◽  
Reference Strain ◽  
Invertible Element ◽  
Matrix Proteins ◽  
Phase Variable ◽  
Content Type ◽  
Third Phase ◽  
Strain Cft073

ABSTRACTUropathogenicEscherichia coli(UPEC) is the most common etiologic agent of uncomplicated urinary tract infection (UTI). An important mechanism of gene regulation in UPEC is phase variation that involves inversion of a promoter-containing DNA element via enzymatic activity of tyrosine recombinases, resulting in biphasic, ON or OFF expression of target genes. The UPEC reference strain CFT073 has five tyrosine site-specific recombinases that function at two previously characterized promoter inversion systems,fimSandhyxS. Three of the five recombinases are located proximally to their cognate target elements, which is typical of promoter inversion systems. The genes for the other two recombinases, IpuA and IpuB, are located distal from these sites. Here, we identified and characterized a third phase-variable invertible element in CFT073,ipuS, located proximal toipuAandipuB. The inversion ofipuSis catalyzed by four of the five CFT073 recombinases. Orientation of the element drives transcription of a two-gene operon containingipuR, a predicted LuxR-type regulator, andupaE, a predicted autotransporter. We show that the predicted autotransporter UpaE is surface located and facilitates biofilm formation as well as adhesion to extracellular matrix proteins in a K-12 recombinant background. Consistent with this phenotype, theipuSON condition in CFT073 results in defective swimming motility, increased adherence to human kidney epithelial cells, and a positive competitive kidney colonization advantage in experimental mouse UTIs. Overall, the identification of a third phase switch in UPEC that is regulated by a shared set of recombinases describes a complex phase-variable virulence network in UPEC.IMPORTANCEUropathogenicEscherichia coli(UPEC) is the most common cause of urinary tract infection (UTI). ON versus OFF phase switching by inversion of small DNA elements at two chromosome sites in UPEC regulates the expression of important virulence factors, including the type 1 fimbria adhesion organelle. In this report, we describe a third invertible element,ipuS, in the UPEC reference strain CFT073. The inversion ofipuScontrols the phase-variable expression ofupaE, an autotransporter gene that encodes a surface protein involved in adherence to extracellular matrix proteins and colonization of the kidneys in a murine model of UTI.

scholarly journals Identification and characterization of a phase-variable element that regulates the autotransporter UpaE in uropathogenicEscherichia coli

2018 ◽  
Author(s):  
E.J. Battaglioli ◽  
K.G.K Goh ◽  
T. S. Atruksang ◽  
K. Schwartz ◽  
M. A. Schembri ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Extracellular Matrix ◽  
Urinary Tract Infection ◽  
Urinary Tract ◽  
Reference Strain ◽  
Invertible Element ◽  
Matrix Proteins ◽  
Phase Variable ◽  
Third Phase ◽  
Strain Cft073

AbstractUropathogenicEscherichia coli(UPEC) are the most common etiological agent of uncomplicated urinary tract infection (UTI). An important mechanism of gene regulation in UPEC is phase variation that involves inversion of a promoter-containing DNA element via enzymatic activity of tyrosine recombinases, resulting in biphasic, ON or OFF expression of target genes. The UPEC reference strain CFT073 has five tyrosine site-specific recombinases that function at two previously characterized promoter inversion systems,fimSandhyxS. Three of the five recombinases are located proximally to their cognate target elements, which is typical of promoter inversion systems. The genes for the other two recombinases, IpuA and IpuB are located distal from these sites. Here, we identified and characterized a third phase variable invertible element in CFT073,ipuSlocated proximal toipuAandipuB. The inversion ofipuSis catalyzed by four of the five CFT073 recombinases. Orientation of the element drives transcription of a two-gene operon containingipuR, a predicted LuxR-type regulator, andupaE, a predicted autotransporter. We show that the predicted autotransporter UpaE is surface-located and facilitates biofilm formation as well as adhesion to extracellular matrix proteins in a K-12 recombinant background. Consistent with this phenotype, theipuSON condition in CFT073 results in defective swimming motility, increased adherence to human kidney epithelial cells, and a positive competitive kidney colonization advantage in experimental mouse UTI infections. Overall, the identification of a third phase-switch in UPEC that is regulated by a shared set of recombinases describes a complex phase-variable virulence network in UPEC.ImportanceUropathogenicEscherichia coli(UPEC) is the most common cause of urinary tract infection (UTI). ON versus OFF phase-switching by inversion of small DNA elements at two chromosome sites in UPEC regulates the expression of important virulence factors, including the type 1 fimbriae adhesion organelle. In this report, we describe a third invertible element,ipuS, in the UPEC reference strain CFT073. The inversion ofipuScontrols the phase variable expression ofupaE, an autotransporter gene that encodes a surface protein involved in adherence to extracellular matrix proteins and colonization of the kidneys in a murine model of UTI.

scholarly journals Long Polar Fimbriae of Enterohemorrhagic Escherichia coli O157:H7 Bind to Extracellular Matrix Proteins

2011 ◽  
Vol 79 (9) ◽  
pp. 3744-3750 ◽  
Author(s):  
Mauricio J. Farfan ◽  
Lidia Cantero ◽  
Roberto Vidal ◽  
Douglas J. Botkin ◽  
Alfredo G. Torres
Keyword(s):  
Escherichia Coli ◽  
Extracellular Matrix ◽  
Enterohemorrhagic Escherichia Coli ◽  
Matrix Proteins ◽  
Intestinal Cells ◽  
T84 Cells ◽  
Content Type ◽  
Fimbrial Subunit ◽  
Ecm Proteins

ABSTRACTAdherence to intestinal cells is a key process in infection caused by enterohemorrhagicEscherichia coli(EHEC). Several adhesion factors that mediate the binding of EHEC to intestinal cells have been described, but the receptors involved in their recognition are not fully characterized. Extracellular matrix (ECM) proteins might act as receptors involved in the recognition of enteric pathogens, including EHEC. In this study, we sought to characterize the binding of EHEC O157:H7 to ECM proteins commonly present in the intestine. We found that EHEC prototype strains as well as other clinical isolates adhered more abundantly to surfaces coated with fibronectin, laminin, and collagen IV. Further characterization of this phenotype, by using antiserum raised against the LpfA1 putative major fimbrial subunit and by addition of mannose, showed that a reduced binding of EHEC to ECM proteins was observed in a long polar fimbria (lpf) mutant. We also found that the two regulators, H-NS and Ler, had an effect in EHEC Lpf-mediated binding to ECM, supporting the roles of these tightly regulated fimbriae as adherence factors. Purified Lpf major subunit bound to all of the ECM proteins tested. Finally, increased bacterial adherence was observed when T84 cells, preincubated with ECM proteins, were infected with EHEC. Taken together, these findings suggest that the interaction of Lpf and ECM proteins contributes to the EHEC colonization of the gastrointestinal tract.

scholarly journals Respiratory Heterogeneity Shapes Biofilm Formation and Host Colonization in Uropathogenic Escherichia coli

mBio ◽  
2019 ◽  
Vol 10 (2) ◽  
Author(s):  
Connor J. Beebout ◽  
Allison R. Eberly ◽  
Sabrina H. Werby ◽  
Seth A. Reasoner ◽  
John R. Brannon ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Extracellular Matrix ◽  
Bacterial Communities ◽  
Spatial Organization ◽  
Bet Hedging ◽  
Content Type ◽  
Dna Organization ◽  
Quinol Oxidases ◽  
Biofilm Development ◽  
Biofilm Community

ABSTRACT Biofilms are multicellular bacterial communities encased in a self-secreted extracellular matrix comprised of polysaccharides, proteinaceous fibers, and DNA. Organization of these components lends spatial organization to the biofilm community such that biofilm residents can benefit from the production of common goods while being protected from exogenous insults. Spatial organization is driven by the presence of chemical gradients, such as oxygen. Here we show that two quinol oxidases found in Escherichia coli and other bacteria organize along the biofilm oxygen gradient and that this spatially coordinated expression controls architectural integrity. Cytochrome bd, a high-affinity quinol oxidase required for aerobic respiration under hypoxic conditions, is the most abundantly expressed respiratory complex in the biofilm community. Depletion of the cytochrome bd-expressing subpopulation compromises biofilm complexity by reducing the abundance of secreted extracellular matrix as well as increasing cellular sensitivity to exogenous stresses. Interrogation of the distribution of quinol oxidases in the planktonic state revealed that ∼15% of the population expresses cytochrome bd at atmospheric oxygen concentration, and this population dominates during acute urinary tract infection. These data point toward a bet-hedging mechanism in which heterogeneous expression of respiratory complexes ensures respiratory plasticity of E. coli across diverse host niches. IMPORTANCE Biofilms are multicellular bacterial communities encased in a self-secreted extracellular matrix comprised of polysaccharides, proteinaceous fibers, and DNA. Organization of these components lends spatial organization in the biofilm community. Here we demonstrate that oxygen gradients in uropathogenic Escherichia coli (UPEC) biofilms lead to spatially distinct expression programs for quinol oxidases—components of the terminal electron transport chain. Our studies reveal that the cytochrome bd-expressing subpopulation is critical for biofilm development and matrix production. In addition, we show that quinol oxidases are heterogeneously expressed in planktonic populations and that this respiratory heterogeneity provides a fitness advantage during infection. These studies define the contributions of quinol oxidases to biofilm physiology and suggest the presence of respiratory bet-hedging behavior in UPEC.

scholarly journals Complete Genome Sequences of Seven Uropathogenic Escherichia coli Strains Isolated from Postmenopausal Women with Recurrent Urinary Tract Infection

2020 ◽  
Vol 9 (33) ◽  
Author(s):  
Belle M. Sharon ◽  
Amber Nguyen ◽  
Amanda P. Arute ◽  
Neha V. Hulyalkar ◽  
Vivian H. Nguyen ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Urinary Tract Infection ◽  
Urinary Tract ◽  
Tract Infection ◽  
Postmenopausal Women ◽  
Complete Genome ◽  
Recurrent Urinary Tract Infection ◽  
Uropathogenic Escherichia Coli ◽  
Genome Sequences ◽  
Content Type

ABSTRACT Uropathogenic Escherichia coli (UPEC) is the most common cause of urinary tract infection (UTI). This disease disproportionately affects women and frequently develops into recurrent UTI (rUTI) in postmenopausal women. Here, we report the complete genome sequences of seven UPEC isolates obtained from the urine of postmenopausal women with rUTI.

scholarly journals Draft Genome Sequence of an Escherichia coli Sequence Type 420 Isolate from a Patient with Urinary Tract Infection in Northern California

2020 ◽  
Vol 9 (23) ◽  
Author(s):  
Yusuke Matsui ◽  
Yuan Hu ◽  
Nicole J. Tarlton ◽  
Lee W. Riley
Keyword(s):  
Escherichia Coli ◽  
Urinary Tract Infection ◽  
Urinary Tract ◽  
Tract Infection ◽  
Genome Sequence ◽  
Draft Genome ◽  
Sequence Type ◽  
Draft Genome Sequence ◽  
Northern California ◽  
Content Type

ABSTRACT The genome sequence of a uropathogenic Escherichia coli sequence type 420 strain isolated from a patient with urinary tract infection in northern California is described here. The draft genome sequence includes a 4.8-Mb chromosome, accompanied by a 114-kb plasmid containing IncFIB/IncFII/Col156 and a 35-kb plasmid containing IncN3.

scholarly journals The Catabolite Repressor Protein-Cyclic AMP Complex RegulatescsgDand Biofilm Formation in Uropathogenic Escherichia coli

2016 ◽  
Vol 198 (24) ◽  
pp. 3329-3334 ◽  
Author(s):  
David A. Hufnagel ◽  
Margery L. Evans ◽  
Sarah E. Greene ◽  
Jerome S. Pinkner ◽  
Scott J. Hultgren ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Extracellular Matrix ◽  
Cyclic Amp ◽  
Intergenic Region ◽  
Biofilm Formation ◽  
Cellulose Production ◽  
Content Type ◽  
E Coli ◽  
Link Type ◽  
K1 Capsule

ABSTRACTThe extracellular matrix protectsEscherichia colifrom immune cells, oxidative stress, predation, and other environmental stresses. Production of theE. coliextracellular matrix is regulated by transcription factors that are tuned to environmental conditions. The biofilm master regulator protein CsgD upregulates curli and cellulose, the two major polymers in the extracellular matrix of uropathogenicE. coli(UPEC) biofilms. We found that cyclic AMP (cAMP) regulates curli, cellulose, and UPEC biofilms throughcsgD. The alarmone cAMP is produced by adenylate cyclase (CyaA), and deletion ofcyaAresulted in reduced extracellular matrix production and biofilm formation. Thecataboliterepressorprotein (CRP) positively regulatedcsgDtranscription, leading to curli and cellulose production in the UPEC isolate, UTI89. Glucose, a known inhibitor of CyaA activity, blocked extracellular matrix formation when added to the growth medium. The mutant strains ΔcyaAand Δcrpdid not produce rugose biofilms, pellicles, curli, cellulose, or CsgD. Three putative CRP binding sites were identified within thecsgD-csgBintergenic region, and purified CRP could gel shift thecsgD-csgBintergenic region. Additionally, we found that CRP binded upstream ofkpsMT, which encodes machinery for K1 capsule production. Together our work shows that cAMP and CRP influenceE. colibiofilms through transcriptional regulation ofcsgD.IMPORTANCEThecataboliterepressorprotein (CRP)-cyclic AMP (cAMP) complex influences the transcription of ∼7% of genes on theEscherichia colichromosome (D. Zheng, C. Constantinidou, J. L. Hobman, and S. D. Minchin, Nucleic Acids Res 32:5874–5893, 2004,https://dx.doi.org/10.1093/nar/gkh908). Glucose inhibitsE. colibiofilm formation, and ΔcyaAand Δcrpmutants show impaired biofilm formation (D. W. Jackson, J.W. Simecka, and T. Romeo, J Bacteriol 184:3406–3410, 2002,https://dx.doi.org/10.1128/JB.184.12.3406-3410.2002). We determined that the cAMP-CRP complex regulates curli and cellulose production and the formation of rugose and pellicle biofilms throughcsgD. Additionally, we propose that cAMP may work as a signaling compound for uropathogenicE. coli(UPEC) to transition from the bladder lumen to inside epithelial cells for intracellular bacterial community formation through K1 capsule regulation.

scholarly journals PhoU enhances the ability of extraintestinal pathogenic Escherichia coli strain CFT073 to colonize the murine urinary tract

Microbiology ◽  
2006 ◽  
Vol 152 (1) ◽  
pp. 153-160 ◽  
Author(s):  
Eric L. Buckles ◽  
Xiaolin Wang ◽  
C. Virginia Lockatell ◽  
David E. Johnson ◽  
Michael S. Donnenberg
Keyword(s):  
Escherichia Coli ◽  
Urinary Tract Infection ◽  
Urinary Tract ◽  
Tract Infection ◽  
Mutant Strain ◽  
Human Urine ◽  
Type Strain ◽  
Wild Type Strain ◽  
Wild Type ◽  
Strain Cft073

The phoU gene is the last cistron in the pstSCAB–phoU operon and functions as a negative regulator of the Pho regulon. The authors previously identified a phoU mutant of extraintestinal pathogenic Escherichia coli strain CFT073 and demonstrated that this mutant was attenuated for survival in the murine model of ascending urinary tract infection. It is hypothesized that the PhoU protein might serve as a urovirulence factor by indirectly affecting the expression of virulence-related genes. In this study, the phoU mutant was further characterized and PhoU was confirmed as a virulence factor. Western blot analysis demonstrated that insertion of the transposon in the phoU gene disrupted the expression of PhoU. The phoU mutant had derepressed alkaline phosphatase activity under phosphate-excess and -limiting conditions. In single-challenge murine ascending urinary tract infection experiments, quantitative cultures of urine, bladder and kidney revealed no significant differences between the phoU mutant strain and the wild-type strain CFT073. However, in competitive colonization experiments, the phoU mutant strain was significantly out-competed by the wild-type strain in the kidneys and urine and recovered in lower amount in the bladder. Complementation of the phoU mutant with a plasmid containing the wild-type phoU gene restored the expression of PhoU and alkaline phosphate activity to wild-type levels and no significant difference in colonization was observed between the phoU mutant containing the complementing plasmid and wild-type in competitive colonization experiments. In human urine, the phoU mutant and wild-type grew comparably when inoculated independently, indicating that the attenuation observed was not due to a general growth defect. However, as observed in vivo, the wild-type out-competed the phoU mutant in competition growth experiments in human urine. These data indicate that PhoU contributes to efficient colonization of the murine urinary tract and add PhoU to a short list of confirmed urovirulence factors.

scholarly journals Inhibition of Escherichia coli CFT073fliCExpression and Motility by Cranberry Materials

2011 ◽  
Vol 77 (19) ◽  
pp. 6852-6857 ◽  
Author(s):  
Gabriela Hidalgo ◽  
Michelle Chan ◽  
Nathalie Tufenkji
Keyword(s):  
Escherichia Coli ◽  
Urinary Tract ◽  
Upper Urinary Tract ◽  
Upec Strain ◽  
Microscopy Imaging ◽  
Content Type ◽  
Reverse Transcription Pcr ◽  
Strain Cft073 ◽  
Tract Infections

ABSTRACTIn humans, uropathogenicEscherichia coli(UPEC) is the most common etiological agent of uncomplicated urinary tract infections (UTIs). Cranberry extracts have been linked to the prevention of UTIs for over a century; however, a mechanistic understanding of the way in which cranberry derivatives prevent bacterial infection is still lacking. In this study, we used afliC-luxreporter as well as quantitative reverse transcription-PCR to demonstrate that when UPEC strain CFT073 was grown or exposed to dehydrated, crushed cranberries or to purified cranberry-derived proanthocyanidins (cPACs), expression of the flagellin gene (fliC) was inhibited. In agreement with these results, transmission electron microscopy imaging of bacteria grown in the presence of cranberry materials revealed fewer flagella than those in bacteria grown under control conditions. Furthermore, we showed that swimming and swarming motilities were hindered when bacteria were grown in the presence of the cranberry compounds. Because flagellum-mediated motility has been suggested to enable UPEC to disseminate to the upper urinary tract, we propose that inhibition of flagellum-mediated motility might be a key mechanism by which cPACs prevent UTIs. This is the first report to show that cranberry compounds inhibit UPEC motility via downregulation of thefliCgene. Further studies are required to establish whether these inhibitors play a rolein vivo.

scholarly journals Characterization of CIA-1, an Ambler Class A Extended-Spectrum β-Lactamase from Chryseobacterium indologenes

2011 ◽  
Vol 56 (1) ◽  
pp. 588-590 ◽  
Author(s):  
Takehisa Matsumoto ◽  
Mika Nagata ◽  
Nau Ishimine ◽  
Kenji Kawasaki ◽  
Kazuyoshi Yamauchi ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Reference Strain ◽  
Content Type ◽  
Chryseobacterium Indologenes ◽  
Esbl Gene ◽  
Class A ◽  
Extended Spectrum ◽  
Class B ◽  
Lactamase Gene

ABSTRACTAn Ambler class A β-lactamase gene,blaCIA-1, was cloned from the reference strainChryseobacterium indologenesATCC 29897 and expressed inEscherichia coliBL21. TheblaCIA-1gene encodes a novel extended-spectrum β-lactamase (ESBL) that shared 68% and 60% identities with the CGA-1 and CME-1 β-lactamases, respectively.blaCIA-1-like genes were detected from clinical isolates. In addition to the metallo-β-lactamase IND of Ambler class B,C. indologeneshas a class A ESBL gene,blaCIA-1, located on the chromosome.

scholarly journals Escherichia coli Isolates That Carryvat,fyuA,chuA, andyfcVEfficiently Colonize the Urinary Tract

2012 ◽  
Vol 80 (12) ◽  
pp. 4115-4122 ◽  
Author(s):  
Rachel R. Spurbeck ◽  
Paul C. Dinh ◽  
Seth T. Walk ◽  
Ann E. Stapleton ◽  
Thomas M. Hooton ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Urinary Tract Infection ◽  
Urinary Tract ◽  
Tract Infection ◽  
Virulence Factors ◽  
Neonatal Meningitis ◽  
Content Type ◽  
E Coli ◽  
Core Set ◽  
Encoding Genes

ABSTRACTExtraintestinalEscherichia coli(ExPEC), a heterogeneous group of pathogens, encompasses avian, neonatal meningitis, and uropathogenicE. colistrains. While several virulence factors are associated with ExPEC, there is no core set of virulence factors that can be used to definitively differentiate these pathotypes. Here we describe a multiplex of four virulence factor-encoding genes,yfcV,vat,fyuA, andchuA, highly associated with uropathogenicE. colistrains that can distinguish three groups ofE. coli: diarrheagenic and animal-associatedE. colistrains, human commensal and avian pathogenicE. colistrains, and uropathogenic and neonatal meningitisE. colistrains. Furthermore, human intestinal isolates that encode all four predictor genes express them during exponential growth in human urine and colonize the bladder in the mouse model of ascending urinary tract infection in higher numbers than human commensal strains that do not encode the four predictor genes (P= 0.02), suggesting that the presence of the predictors correlates with uropathogenic potential.

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