scholarly journals The Serine Protease Autotransporters TagB, TagC, and Sha from Extraintestinal Pathogenic Escherichia coli Are Internalized by Human Bladder Epithelial Cells and Cause Actin Cytoskeletal Disruption

2020 ◽  
Vol 21 (9) ◽  
pp. 3047 ◽  
Author(s):  
Pravil Pokharel ◽  
Juan Manuel Díaz ◽  
Hicham Bessaiah ◽  
Sébastien Houle ◽  
Alma Lilián Guerrero-Barrera ◽  
...  
Keyword(s):  
Urinary Tract ◽  
Epithelial Cells ◽  
Serine Protease ◽  
Catalytic Triad ◽  
Host Cells ◽  
Bacterial Cells ◽  
Bladder Epithelium ◽  
Cytopathic Effects ◽  
Important Barrier ◽  
Bladder Cells

TagB, TagC (tandem autotransporter genes B and C), and Sha (Serine-protease hemagglutinin autotransporter) are recently described members of the SPATE (serine protease autotransporters of Enterobacteriaceae) family. These SPATEs can cause cytopathic effects on bladder cells and contribute to urinary tract infection in a mouse model. Bladder epithelial cells form an important barrier in the urinary tract. Some SPATEs produced by pathogenic E. coli are known to breach the bladder epithelium. The capacity of these newly described SPATEs to alter bladder epithelial cells and the role of the serine protease active site were investigated. All three SPATE proteins were internalized by bladder epithelial cells and altered the distribution of actin cytoskeleton. Sha and TagC were also shown to degrade mucin and gelatin respectively. Inactivation of the serine catalytic site in each of these SPATEs did not affect secretion of the SPATEs from bacterial cells, but abrogated entry into epithelial cells, cytotoxicity, and proteolytic activity. Thus, our results show that the serine catalytic triad of these proteins is required for internalization in host cells, actin disruption, and degradation of host substrates such as mucin and gelatin.

scholarly journals Dynamic localisation of DamX regulates bacterial filamentation and division during UPEC dispersal from host cells

2021 ◽  
Author(s):  
Bill Soderstrom ◽  
Daniel O. Daley ◽  
Iain G. Duggin
Keyword(s):  
Escherichia Coli ◽  
Cell Division ◽  
Stress Response ◽  
Epithelial Cells ◽  
Membrane Fusion ◽  
Host Cells ◽  
Division Rings ◽  
Bladder Cells ◽  
Rod Cell

Uropathogenic Escherichia coli (UPEC) cells can grow into highly filamentous forms during infection of bladder epithelial cells, but this process is poorly understood. Herein we found that some UPEC filaments released from infected bladder cells in vitro grew very rapidly and by more than 100 μm before initiating division, whereas others did not survive, suggesting that filamentation is a stress response that promotes dispersal. The DamX bifunctional division protein, which is essential for UPEC filamentation, was initially non-localized but then assembled at multiple division sites in the filaments prior to division. DamX rings maintained consistent thickness during constriction and remained at the septum until after membrane fusion was completed, like in rod cell division. Our findings suggest a mechanism involving regulated dissipation of DamX, leading to division arrest and filamentation, followed by its reassembly into division rings to promote UPEC dispersal and survival during infection.

scholarly journals Escherichia coli DraE Adhesin-Associated Bacterial Internalization by Epithelial Cells Is Promoted Independently by Decay-Accelerating Factor and Carcinoembryonic Antigen-Related Cell Adhesion Molecule Binding and Does Not Require the DraD Invasin

2008 ◽  
Vol 76 (9) ◽  
pp. 3869-3880 ◽  
Author(s):  
Natalia Korotkova ◽  
Yuliya Yarova-Yarovaya ◽  
Veronika Tchesnokova ◽  
Nina Yazvenko ◽  
Mike A. Carl ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Cell Adhesion ◽  
Epithelial Cells ◽  
Carcinoembryonic Antigen ◽  
Host Cells ◽  
Decay Accelerating Factor ◽  
E Coli ◽  
Related Cell ◽  
Bladder Cells ◽  
Tract Infections

ABSTRACT The Dr family of Escherichia coli adhesins are virulence factors associated with diarrhea and urinary tract infections. Dr fimbriae are comprised of two subunits. DraE/AfaE represents the major structural, antigenic, and adhesive subunit, which recognizes decay-accelerating factor (DAF) and carcinoembryonic antigen (CEA)-related cell adhesion molecules (CEACAMs) CEA, CEACAM1, CEACAM3, and CEACAM6 as binding receptors. The DraD/AfaD subunit caps fimbriae and has been implicated in the entry of Dr-fimbriated E. coli into host cells. In this study, we demonstrate that DAF or CEACAM receptors independently promote DraE-mediated internalization of E. coli by CHO cell transfectants expressing these receptors. We also found that DraE-positive recombinant bacteria adhere to and are internalized by primary human bladder epithelial cells which express DAF and CEACAMs. DraE-mediated bacterial internalization by bladder cells was inhibited by agents which disrupt lipid rafts, microtubules, and phosphatidylinositol 3-kinase (PI3K) activity. Immunofluorescence confocal microscopic examination of epithelial cells detected considerable recruitment of caveolin, β1 integrin, phosphorylated ezrin, phosphorylated PI3K, and tubulin, but not F-actin, by cell-associated bacteria. Finally, we demonstrate that the DraD subunit, previously implicated as an “invasin,” is not required for β1 integrin recruitment or bacterial internalization.

scholarly journals Establishment of a Persistent Escherichia coli Reservoir during the Acute Phase of a Bladder Infection

2001 ◽  
Vol 69 (7) ◽  
pp. 4572-4579 ◽  
Author(s):  
Matthew A. Mulvey ◽  
Joel D. Schilling ◽  
Scott J. Hultgren
Keyword(s):  
Escherichia Coli ◽  
Urinary Tract ◽  
Epithelial Cells ◽  
Urinary Tract Infections ◽  
Bacterial Attachment ◽  
Bacterial Invasion ◽  
E Coli ◽  
Bladder Cells ◽  
Tract Infections

ABSTRACT The vast majority of urinary tract infections are caused by strains of uropathogenic Escherichia coli that encode filamentous adhesive organelles called type 1 pili. These structures mediate both bacterial attachment to and invasion of bladder epithelial cells. However, the mechanism by which type 1 pilus-mediated bacterial invasion contributes to the pathogenesis of a urinary tract infection is unknown. Here we show that type 1-piliated uropathogens can invade the superficial epithelial cells that line the lumenal surface of the bladder and subsequently replicate, forming massive foci of intracellular E. coli termed bacterial factories. In response to infection, superficial bladder cells exfoliate and are removed with the flow of urine. To avoid clearance by exfoliation, intracellular uropathogens can reemerge and eventually establish a persistent, quiescent bacterial reservoir within the bladder mucosa that may serve as a source for recurrent acute infections. These observations suggest that urinary tract infections are more chronic and invasive than generally assumed.

scholarly journals The Potential Virulence Factors ofProvidencia stuartii: Motility, Adherence, and Invasion

2018 ◽  
Vol 2018 ◽  
pp. 1-8 ◽  
Author(s):  
Naziia Kurmasheva ◽  
Vyacheslav Vorobiev ◽  
Margarita Sharipova ◽  
Tatyana Efremova ◽  
Ayslu Mardanova
Keyword(s):  
Epithelial Cells ◽  
Nk Cells ◽  
Morphological Changes ◽  
Viscous Medium ◽  
Host Cells ◽  
Bacterial Cells ◽  
M Cell ◽  
Infectious Dose ◽  
Tract Infections

Providencia stuartiiis the most commonProvidenciaspecies capable of causing human infections. CurrentlyP. stuartiiis involved in high incidence of urinary tract infections in catheterized patients. The ability of bacteria to swarm on semisolid (viscous) surfaces and adhere to and invade host cells determines the specificity of the disease pathogenesis and its therapy. In the present study we demonstrated morphological changes ofP. stuartiiNK cells during migration on the viscous medium and discussed adhesive and invasive properties utilizing the HeLa-M cell line as a host model. To visualize the interaction ofP. stuartiiNK bacterial cells with eukaryotic cellsin vitroscanning electron and confocal microscopy were performed. We found that bacteriaP. stuartiiNK are able to adhere to and invade HeLa-M epithelial cells and these properties depend on the age of bacterial culture. Also, to invade the host cells the infectious dose of the bacteria is essential. The microphotographs indicate that after incubation of bacterialP. stuartiiNK cells together with epithelial cells the bacterial cells both were adhered onto and invaded into the host cells.

scholarly journals Bacterial Lysis Liberates the Neutrophil Migration Suppressor YbcL from the Periplasm of Uropathogenic Escherichia coli

2014 ◽  
Vol 82 (12) ◽  
pp. 4921-4930 ◽  
Author(s):  
Megan E. Lau ◽  
Elizabeth S. Danka ◽  
Kristin M. Tiemann ◽  
David A. Hunstad
Keyword(s):  
Escherichia Coli ◽  
Urinary Tract ◽  
Epithelial Cells ◽  
Neutrophil Migration ◽  
Outer Membrane Vesicles ◽  
Bacterial Invasion ◽  
Bladder Epithelium ◽  
Content Type

ABSTRACTUropathogenicEscherichia coli(UPEC) modulates aspects of the innate immune response during urinary tract infection to facilitate bacterial invasion of the bladder epithelium, a requirement for the propagation of infection. For example, UPEC-encoded YbcL suppresses the traversal of bladder epithelia by neutrophils in both anin vitromodel and anin vivomurine cystitis model. The suppressive activity of YbcL requires liberation from the bacterial periplasm, though the mechanism of release is undefined. Here we present findings on the site of action of YbcL and demonstrate a novel mode of secretion for a UPEC exoprotein. Suppression of neutrophil migration by purified YbcLUTI, encoded by cystitis isolate UTI89, required the presence of a uroepithelial layer; YbcLUTIdid not inhibit neutrophil chemotaxis directly. YbcLUTIwas released to a greater extent during UPEC infection of uroepithelial cells than during that of neutrophils. Release of YbcLUTIwas maximal when UPEC and bladder epithelial cells were in close proximity. Established modes of secretion, including outer membrane vesicles, the type II secretion system, and the type IV pilus, were dispensable for YbcLUTIrelease from UPEC. Instead, YbcLUTIwas liberated during bacterial death, which was augmented upon exposure to bladder epithelial cells, as confirmed by detection of bacterial cytoplasmic proteins and DNA in the supernatant and enumeration of bacteria with compromised membranes. As YbcLUTIacts on the uroepithelium to attenuate neutrophil migration, this mode of release may represent a type of altruistic cooperation within a UPEC population during colonization of the urinary tract.

scholarly journals In vivo replacement of damaged bladder urothelium by Wolffian duct epithelial cells

2018 ◽  
Vol 115 (33) ◽  
pp. 8394-8399 ◽  
Author(s):  
Diya B. Joseph ◽  
Anoop S. Chandrashekar ◽  
Lisa L. Abler ◽  
Li-Fang Chu ◽  
James A. Thomson ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Dna Methyltransferase ◽  
Wolffian Duct ◽  
Bladder Epithelium ◽  
Bladder Urothelium ◽  
Keratin 5 ◽  
Bladder Regeneration ◽  
Bladder Cells ◽  
And Function

The bladder’s remarkable regenerative capacity had been thought to derive exclusively from its own progenitors. While examining consequences of DNA methyltransferase 1 (Dnmt1) inactivation in mouse embryonic bladder epithelium, we made the surprising discovery that Wolffian duct epithelial cells can support bladder regeneration. Conditional Dnmt1 inactivation in mouse urethral and bladder epithelium triggers widespread apoptosis, depletes basal and intermediate bladder cells, and disrupts uroplakin protein expression. These events coincide with Wolffian duct epithelial cell recruitment into Dnmt1 mutant urethra and bladder where they are reprogrammed to express bladder markers, including FOXA1, keratin 5, P63, and uroplakin. This is evidence that Wolffian duct epithelial cells are summoned in vivo to replace damaged bladder epithelium and function as a reservoir of cells for bladder regeneration.

scholarly journals Distinct Morphological Fates of Uropathogenic Escherichia coli Intracellular Bacterial Communities: Dependency on Urine Composition and pH

2020 ◽  
Vol 88 (9) ◽  
Author(s):  
Gregory Iosifidis ◽  
Iain G. Duggin
Keyword(s):  
Escherichia Coli ◽  
Urinary Tract ◽  
Epithelial Cells ◽  
Human Urine ◽  
Bacterial Communities ◽  
Acute Infection ◽  
Infection Cycle ◽  
Bladder Cells ◽  
Tract Infections ◽  
Intracellular Bacterial Communities

ABSTRACT Uropathogenic Escherichia coli (UPEC) is the leading cause of urinary tract infections. These bacteria undertake a multistage infection cycle involving invasion of and proliferation within urinary tract epithelial cells, leading to the rupture of the host cell and dispersal of the bacteria, some of which have a highly filamentous morphology. Here, we established a microfluidics-based model of UPEC infection of immortalized human bladder epithelial cells that recapitulates the main stages of bacterial morphological changes during the acute infection cycle in vivo and allows the development and fate of individual cells to be monitored in real time by fluorescence microscopy. The UPEC-infected bladder cells remained alive and mobile in nonconfluent monolayers during the development of intracellular bacterial communities (IBCs). Switching from a flow of growth medium to human urine resulted in immobilization of both uninfected and infected bladder cells. Some IBCs continued to develop and then released many highly filamentous bacteria via an extrusion-like process, whereas other IBCs showed strong UPEC proliferation, and yet no filamentation was detected. The filamentation response was dependent on the weak acidity of human urine and required component(s) in a low molecular-mass (<3,000 Da) fraction from a mildly dehydrated donor. The developmental fate for bacteria therefore appears to be controlled by multiple factors that act at the level of the whole IBC, suggesting that variable local environments or stochastic differentiation pathways influence IBC developmental fates during infection.

scholarly journals The Serine Protease Motif of EspC from Enteropathogenic Escherichia coli Produces Epithelial Damage by a Mechanism Different from That of Pet Toxin from Enteroaggregative E. coli

2004 ◽  
Vol 72 (6) ◽  
pp. 3609-3621 ◽  
Author(s):  
Fernando Navarro-García ◽  
Adrián Canizalez-Roman ◽  
Bao Quan Sui ◽  
James P. Nataro ◽  
Yenia Azamar
Keyword(s):  
Escherichia Coli ◽  
Epithelial Cells ◽  
Serine Protease ◽  
Protein C ◽  
Conformational Structure ◽  
Cleavage Sites ◽  
E Coli ◽  
Calmodulin Binding ◽  
Cytopathic Effects ◽  
Cytoskeleton Disruption

ABSTRACT EspC (Escherichia coli secreted protein C) of enteropathogenic E. coli (EPEC) shows the three classical domains of the autotransporter proteins and has a conserved serine protease motif belonging to the SPATE (serine protease autotransporters of Enterobacteriaceae) subfamily. EspC and its homolog Pet in enteroaggregative E. coli (EAEC) bear the same sequence within the serine protease motif, and both proteins produce enterotoxic effects, suggesting that like Pet, EspC could be internalized to reach and cleave the calmodulin-binding domain of fodrin, causing actin cytoskeleton disruption. Even though both proteins cause cytoskeleton damage by virtue of their serine protease motifs, the following evidence supports the hypothesis that the mechanisms are different. (i) To obtain similar cytotoxic and cytoskeletal effects, a threefold-higher EspC concentration and a twofold-higher exposure time are needed. (ii) EspC internalization into epithelial cells takes more time (6 h) than Pet internalization (30 min), and the distributions of the two proteins inside the cells are also different. (iii) Both proteins have affinity for fodrin and cleave it, but the cleavage sites are different; EspC produces two cleavages, while Pet produces just one. (iv) EspC does not cause fodrin redistribution within epithelial cells. (v) An EspC serine protease motif mutant, but not a Pet serine protease mutant, competes with EspC by blocking cytoskeletal damage. All these data suggest that the protein conformational structure is very important for the activity of the catalytic site, influencing its interaction with the target protein and its internalization. The differences between these proteins may explain the reduced ability of EspC to cause cytopathic effects. However, these differences may confer a specialized role on EspC in the pathogenesis of EPEC, which is different from that of Pet in EAEC pathogenesis.

scholarly journals An infection-induced RhoB-Beclin 1-Hsp90 complex enhances clearance of uropathogenic Escherichia coli

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Chunhui Miao ◽  
Mingyu Yu ◽  
Geng Pei ◽  
Zhenyi Ma ◽  
Lisong Zhang ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Treatment Strategies ◽  
Uropathogenic Escherichia Coli ◽  
Beclin 1 ◽  
Host Cells ◽  
Infection Model ◽  
Intracellular Bacteria ◽  
Bladder Epithelium ◽  
Binding Residue

AbstractHost cells use several anti-bacterial pathways to defend against pathogens. Here, using a uropathogenic Escherichia coli (UPEC) infection model, we demonstrate that bacterial infection upregulates RhoB, which subsequently promotes intracellular bacteria clearance by inducing LC3 lipidation and autophagosome formation. RhoB binds with Beclin 1 through its residues at 118 to 140 and the Beclin 1 CCD domain, with RhoB Arg133 being the key binding residue. Binding of RhoB to Beclin 1 enhances the Hsp90-Beclin 1 interaction, preventing Beclin 1 degradation. RhoB also directly interacts with Hsp90, maintaining RhoB levels. UPEC infections increase RhoB, Beclin 1 and LC3 levels in bladder epithelium in vivo, whereas Beclin 1 and LC3 levels as well as UPEC clearance are substantially reduced in RhoB+/− and RhoB−/− mice upon infection. We conclude that when stimulated by UPEC infections, host cells promote UPEC clearance through the RhoB-Beclin 1-HSP90 complex, indicating RhoB may be a useful target when developing UPEC treatment strategies.

scholarly journals Cigarette Smoke Stimulates SARS-CoV-2 Internalization by Activating AhR and Increasing ACE2 Expression in Human Gingival Epithelial Cells

2021 ◽  
Vol 22 (14) ◽  
pp. 7669
Author(s):  
Cassio Luiz Coutinho Almeida-da-Silva ◽  
Harmony Matshik Dakafay ◽  
Kaitlyn Liu ◽  
David M. Ojcius
Keyword(s):  
Epithelial Cells ◽  
Oral Cavity ◽  
Cigarette Smoke ◽  
Large Body ◽  
Receptor Expression ◽  
Host Cells ◽  
Dependent Manner ◽  
Gingival Epithelial Cells ◽  
Human Gingival Epithelial Cells ◽  
Oral Cells

A large body of evidence shows the harmful effects of cigarette smoke to oral and systemic health. More recently, a link between smoking and susceptibility to coronavirus disease 2019 (COVID-19) was proposed. COVID-19 is due to infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which uses the receptor ACE2 and the protease TMPRSS2 for entry into host cells, thereby infecting cells of the respiratory tract and the oral cavity. Here, we examined the effects of cigarette smoke on the expression of SARS-CoV-2 receptors and infection in human gingival epithelial cells (GECs). We found that cigarette smoke condensates (CSC) upregulated ACE2 and TMPRSS2 expression in GECs, and that CSC activated aryl hydrocarbon receptor (AhR) signaling in the oral cells. ACE2 was known to mediate SARS-CoV-2 internalization, and we demonstrate that CSC treatment potentiated the internalization of SARS-CoV-2 pseudovirus in GECs in an AhR-dependent manner. AhR depletion using small interference RNA decreased SARS-CoV-2 pseudovirus internalization in CSC-treated GECs compared with control GECs. Our study reveals that cigarette smoke upregulates SARS-CoV-2 receptor expression and infection in oral cells. Understanding the mechanisms involved in SARS-CoV-2 infection in cells of the oral cavity may suggest therapeutic interventions for preventing viral infection and transmission.

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