Clathrin, AP-2, and the NPXY-binding subset of alternate endocytic adaptors facilitate FimH-mediated bacterial invasion of host cells

Eto, Danelle S.; Gordon, Hannah B.; Dhakal, Bijaya K.; Jones, Tiffani A.; Mulvey, Matthew

doi:10.1111/j.1462-5822.2008.01229.x

Cited by 74 publications

(55 citation statements)

References 94 publications

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“…Host cell association and gentamicin protection-based invasion assays were performed as previously described (66,67). Strains used in these assays were grown at 37°C for 48 h in static LB broth to induce expression of type 1 pili.…”

Section: Methodsmentioning

confidence: 99%

The Cpx Stress Response System Potentiates the Fitness and Virulence of Uropathogenic Escherichia coli

et al. 2013

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Strains of uropathogenic Escherichia coli (UPEC) are the primary cause of urinary tract infections, representing one of the most widespread and successful groups of pathogens on the planet. To colonize and persist within the urinary tract, UPEC must be able to sense and respond appropriately to environmental stresses, many of which can compromise the bacterial envelope. The Cpx two-component envelope stress response system is comprised of the inner membrane histidine kinase CpxA, the cytosolic response regulator CpxR, and the periplasmic auxiliary factor CpxP. Here, by using deletion mutants along with mouse and zebrafish infection models, we show that the Cpx system is critical to the fitness and virulence of two reference UPEC strains, the cystitis isolate UTI89 and the urosepsis isolate CFT073. Specifically, deletion of the cpxRA operon impaired the ability of UTI89 to colonize the murine bladder and greatly reduced the virulence of CFT073 during both systemic and localized infections within zebrafish embryos. These defects coincided with diminished host cell invasion by UTI89 and increased sensitivity of both strains to complement-mediated killing and the aminoglycoside antibiotic amikacin. Results obtained with the cpxP deletion mutants were more complicated, indicating variable strain-dependent and niche-specific requirements for this well-conserved auxiliary factor.

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Section: Methodsmentioning

confidence: 99%

The Cpx Stress Response System Potentiates the Fitness and Virulence of Uropathogenic Escherichia coli

et al. 2013

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“…Protein concentrations within the lysates were determined using the BCA reagent system (Pierce), and equivalent protein amounts were resolved by SDS-PAGE and transferred to an Immobilon PVDF-FL membrane (Millipore). Blots were probed using anti-RpoS (Neoclone) and anti-E. coli antibodies (Biodesign International) and visualized using enhanced chemiluminescence as previously described (30). pH stress resistance assays.…”

Section: Methodsmentioning

confidence: 99%

Adenylate Cyclase and the Cyclic AMP Receptor Protein Modulate Stress Resistance and Virulence Capacity of Uropathogenic Escherichia coli

et al. 2013

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In many bacteria, the second messenger cyclic AMP (cAMP) interacts with the transcription factor cAMP receptor protein (CRP), forming active cAMP-CRP complexes that can control a multitude of cellular activities, including expanded carbon source utilization, stress response pathways, and virulence. Here, we assessed the role of cAMP-CRP as a regulator of stress resistance and virulence in uropathogenic Escherichia coli (UPEC), the principal cause of urinary tract infections worldwide. Deletion of genes encoding either CRP or CyaA, the enzyme responsible for cAMP synthesis, attenuates the ability of UPEC to colonize the bladder in a mouse infection model, dependent on intact innate host defenses. UPEC mutants lacking cAMP-CRP grow normally in the presence of glucose but are unable to utilize alternate carbon sources like amino acids, the primary nutrients available to UPEC within the urinary tract. Relative to the wild-type UPEC isolate, the cyaA and crp deletion mutants are sensitive to nitrosative stress and the superoxide generator methyl viologen but remarkably resistant to hydrogen peroxide (H 2 O 2 ) and acid stress. In the mutant strains, H 2 O 2 resistance correlates with elevated catalase activity attributable in part to enhanced translation of the alternate sigma factor RpoS. Acid resistance was promoted by both RpoS-independent and RpoS-dependent mechanisms, including expression of the RpoS-regulated DNA-binding ferritin-like protein Dps. We conclude that balanced input from many cAMP-CRP-responsive elements, including RpoS, is critical to the ability of UPEC to handle the nutrient limitations and severe environmental stresses present within the mammalian urinary tract. U nder homeostatic conditions, the mammalian urinary tract is maintained as a sterile environment through the production of antimicrobial peptides and other toxic compounds, the bulk flow of urine, innate immune cell surveillance mechanisms, and nutrient limitations (1-5). However, select microbial pathogens are capable of colonizing and infecting this normally inhospitable niche. Uropathogenic Escherichia coli (UPEC) is the major cause of urinary tract infections (UTI) worldwide, affecting millions and requiring billions of dollars for diagnosis and treatment annually (6). To overcome host defenses and effectively colonize the urinary tract, UPEC employs a variety of virulence factors and stress resistance mechanisms, including adhesive and motility organelles that mediate attachment to and invasion of host cells, toxins that disarm innate immune responses, and multiple iron-scavenging proteins (1, 7-9). The ability to sense and prioritize the use of limited carbon sources within the nutrient-poor environment of the urinary tract is also likely critical to the success of UPEC, but our understanding of the impact that bacterial metabolic pathways have on the establishment and progression of a UTI is incomplete.Within the urinary tract, UPEC relies largely on the catabolism of small peptides and amino acids for survival and growth (4). U...

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“…The coordinated activation of these host factors drives localized rearrangements of the actin cytoskeleton, leading to the envelopment and internalization of bound UPEC. This is a cholesterol-dependent process that also requires clathrin, the canonical clathrin adaptor AP-2, and at least three alternate endocytic adaptors, Numb, ARH, and Dab2 (19).…”

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confidence: 99%

Uropathogenic Escherichia coli Invades Host Cells via an HDAC6-modulated Microtubule-dependent Pathway

Dhakal

Mulvey

2009

Journal of Biological Chemistry

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Strains of uropathogenic Escherichia coli (UPEC) encode filamentous adhesive organelles called type 1 pili that promote bacterial colonization and invasion of the bladder epithelium. Type 1 pilus-mediated interactions with host receptors, including ␣3␤1 integrin, trigger localized actin rearrangements that lead to internalization of adherent bacteria via a zipper-like mechanism. Here we report that type 1 pilus-mediated bacterial invasion of bladder cells also requires input from host microtubules and histone deacetylase 6 (HDAC6), a cytosolic enzyme that, by deacetylating ␣-tubulin, can alter the stability of microtubules along with the recruitment and directional trafficking of the kinesin-1 motor complex. We found that disruption of microtubules by nocodazole or vinblastine treatment, as well as microtubule stabilization by taxol, inhibited host cell invasion by UPEC, as did silencing of HDAC6 expression or pharmacological inhibition of HDAC6 activity. Invasion did not require two alternate HDAC6 substrates, Hsp90 and cortactin, but was dependent upon the kinesin-1 light chain KLC2 and an upstream activator of HDAC6, aurora A kinase. These results indicate that HDAC6 and microtubules act as vital regulatory elements during the invasion process, possibly via indirect effects on kinesin-1 and associated cargos.

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Clathrin, AP-2, and the NPXY-binding subset of alternate endocytic adaptors facilitate FimH-mediated bacterial invasion of host cells

Cited by 74 publications

References 94 publications

The Cpx Stress Response System Potentiates the Fitness and Virulence of Uropathogenic Escherichia coli

The Cpx Stress Response System Potentiates the Fitness and Virulence of Uropathogenic Escherichia coli

Adenylate Cyclase and the Cyclic AMP Receptor Protein Modulate Stress Resistance and Virulence Capacity of Uropathogenic Escherichia coli

Uropathogenic Escherichia coli Invades Host Cells via an HDAC6-modulated Microtubule-dependent Pathway

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