Escherichia coli Biofilms Have an Organized and Complex Extracellular Matrix Structure

Hung, Chia-Suei; Zhou, Yizhou; Pinkner, Jerome S.; Dodson, Karen W.; Crowley, Jan R.; Heuser, John E.; Chapman, Matthew R.; Hadjifrangiskou, Maria; Henderson, Jeffrey P.; Hultgren, Scott J.

doi:10.1128/mbio.00645-13

Cited by 208 publications

(198 citation statements)

References 67 publications

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“…1E) (39). Like rugose colonies, pellicle biofilms are dependent on both curli and cellulose (40). A ⌬dsbB mutant formed a more robust and wrinkled pellicle than did the WT (Fig.…”

Section: Disulfide Bonding Mutants Form Hyperbiofilmsmentioning

confidence: 99%

The Disulfide Bonding System Suppresses CsgD-Independent Cellulose Production in Escherichia coli

2014

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The bacterial extracellular matrix encases cells and protects them from host-related and environmental insults. The Escherichia coli master biofilm regulator CsgD is required for the production of the matrix components curli and cellulose. CsgD activates the diguanylate cyclase AdrA, which in turn stimulates cellulose production through cyclic di-GMP (c-di-GMP). Here, we identified and characterized a CsgD-and AdrA-independent cellulose production pathway that was maximally active when cultures were grown under reducing conditions or when the disulfide bonding system (DSB) was compromised. The CsgD-independent cellulose activation pathway was dependent on a second diguanylate cyclase, called YfiN. c-di-GMP production by YfiN was repressed by the periplasmic protein YfiR, and deletion of yfiR promoted CsgD-independent cellulose production. Conversely, when YfiR was overexpressed, cellulose production was decreased. Finally, we found that YfiR was oxidized by DsbA and that intraprotein YfiR disulfide bonds stabilized YfiR in the periplasm. Altogether, we showed that reducing conditions and mutations in the DSB system caused hyperactivation of YfiN and subsequent CsgD-independent cellulose production.

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“…1E) (39). Like rugose colonies, pellicle biofilms are dependent on both curli and cellulose (40). A ⌬dsbB mutant formed a more robust and wrinkled pellicle than did the WT (Fig.…”

Section: Disulfide Bonding Mutants Form Hyperbiofilmsmentioning

confidence: 99%

The Disulfide Bonding System Suppresses CsgD-Independent Cellulose Production in Escherichia coli

2014

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“…coli pellicles are curli-and cellulose-dependent biofilms that form at the air-liquid interface of static cultures (10,27). To assess whether glucose inhibits pellicle biofilm formation, UTI89 was grown in liquid cultures in the absence or presence of glucose for 48 h at 26°C.…”

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confidence: 99%

The Catabolite Repressor Protein-Cyclic AMP Complex Regulates csgD and Biofilm Formation in Uropathogenic Escherichia coli

et al. 2016

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The extracellular matrix protects Escherichia coli from immune cells, oxidative stress, predation, and other environmental stresses. Production of the E. coli extracellular 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 uropathogenic E. coli (UPEC) biofilms. We found that cyclic AMP (cAMP) regulates curli, cellulose, and UPEC biofilms through csgD. The alarmone cAMP is produced by adenylate cyclase (CyaA), and deletion of cyaA resulted in reduced extracellular matrix production and biofilm formation. The catabolite repressor protein (CRP) positively regulated csgD transcription, 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 ⌬cyaA and ⌬crp did not produce rugose biofilms, pellicles, curli, cellulose, or CsgD. Three putative CRP binding sites were identified within the csgD-csgB intergenic region, and purified CRP could gel shift the csgD-csgB intergenic region. Additionally, we found that CRP binded upstream of kpsMT, which encodes machinery for K1 capsule production. Together our work shows that cAMP and CRP influence E. coli biofilms through transcriptional regulation of csgD. IMPORTANCE The catabolite repressor protein (CRP)-cyclic AMP (cAMP) complex influences the transcription of ϳ7% of genes on the Escherichia coli is a multifaceted Gram-negative bacterium that is equipped for survival in disparate environments ranging from the gastrointestinal and urinary tracts to waterways and soil (1). Growth of E. coli can occur with either planktonic or biofilm lifestyles, the transition between which requires a highly organized shift in gene expression. The biofilm lifestyle is characterized by adherence, slow growth, resistance to environmental insults, and production of an extracellular matrix (ECM) (2). CsgD is the major biofilm regulator of E. coli and of many other bacteria of the Enterobacteriaceae family (3-5), and it promotes the production of two E. coli surface appendages, the amyloid fiber curli and the fibrous glycan chain cellulose (2). CsgD represses the transcription of motility-associated genes, while mediating self and surface attachment (6-8).E. coli is found in ϳ80% of urinary tract infections (UTI), more than any other causative agent, and has the ability to thrive in the bladder and urinary tract environments (9). During UTI, uropathogenic E. coli (UPEC) uses various means to evade and colonize the host. Curli and other factors function in establishing an initial bacterial titer (10). Once in the bladder, E. coli utilizes type 1 pili to invade bladder epithelial cells (11). E. coli bacteria then form a biofilm-like structure known as an intracellular bacterial community (IBC), and cells are coated by the ECM component K1 for protection from host cell recognition ...

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“…Escherichia coli, Salmonella spp., and many other Enterobacteriaceae produce functional amyloid fibers, called curli, on their surfaces (3)(4)(5)(6) that are involved in cell adherence, invasion, host colonization, and biofilm formation (6)(7)(8)(9)(10)(11). Curli fibers make up the primary proteinaceous component of the extracellular matrix in pellicle biofilm, a subset of biofilms formed by the cystitis uropathogenic E. coli isolate UTI89 (12). The formation of biofilms represents a common strategy by which bacteria resist mechanical and chemical clearance mechanisms of the host (13)(14)(15).…”

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confidence: 99%

Solution NMR structure of CsgE: Structural insights into a chaperone and regulator protein important for functional amyloid formation

Krezel

Cusumano

Pinkner

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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Curli, consisting primarily of major structural subunit CsgA, are functional amyloids produced on the surface of Escherichia coli, as well as many other enteric bacteria, and are involved in cell colonization and biofilm formation. CsgE is a periplasmic accessory protein that plays a crucial role in curli biogenesis. CsgE binds to both CsgA and the nonameric pore protein CsgG. The CsgG-CsgE complex is the curli secretion channel and is essential for the formation of the curli fibril in vivo. To better understand the role of CsgE in curli formation, we have determined the solution NMR structure of a double mutant of CsgE (W48A/F79A) that appears to be similar to the wild-type (WT) protein in overall structure and function but does not form mixed oligomers at NMR concentrations similar to the WT. The well-converged structure of this mutant has a core scaffold composed of a layer of two α-helices and a layer of three-stranded antiparallel β-sheet with flexible N and C termini. The structure of CsgE fits well into the cryoelectron microscopy density map of the CsgG-CsgE complex. We highlight a striking feature of the electrostatic potential surface in CsgE structure and present an assembly model of the CsgG-CsgE complex. We suggest a structural mechanism of the interaction between CsgE and CsgA. Understanding curli formation can provide the information necessary to develop treatments and therapeutic agents for biofilm-related infections and may benefit the prevention and treatment of amyloid diseases. CsgE could establish a paradigm for the regulation of amyloidogenesis because of its unique role in curli formation.biofilm formation | intrinsically disordered protein | aggregation | protein-protein interaction | CsgG

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Escherichia coli Biofilms Have an Organized and Complex Extracellular Matrix Structure

Cited by 208 publications

References 67 publications

The Disulfide Bonding System Suppresses CsgD-Independent Cellulose Production in Escherichia coli

The Disulfide Bonding System Suppresses CsgD-Independent Cellulose Production in Escherichia coli

The Catabolite Repressor Protein-Cyclic AMP Complex Regulates csgD and Biofilm Formation in Uropathogenic Escherichia coli

Solution NMR structure of CsgE: Structural insights into a chaperone and regulator protein important for functional amyloid formation

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