The Phosphoenolpyruvate Phosphotransferase System: as Important for Biofilm Formation by Vibrio cholerae as It Is for Metabolism in Escherichia coli

AbstractBiofilm formation protects bacteria from stresses including antibiotics and host immune responses. Carbon sources can modulate biofilm formation and host colonization in Vibrio cholerae, but the underlying mechanisms remain unclear. Here, we show that EIIAGlc, a component of the phosphoenolpyruvate (PEP):carbohydrate phosphotransferase system (PTS), regulates the intracellular concentration of the cyclic dinucleotide c-di-GMP, and thus biofilm formation. The availability of preferred sugars such as glucose affects EIIAGlc phosphorylation state, which in turn modulates the interaction of EIIAGlc with a c-di-GMP phosphodiesterase (hereafter referred to as PdeS). In a Drosophila model of V. cholerae infection, sugars in the host diet regulate gut colonization in a manner dependent on the PdeS-EIIAGlc interaction. Our results shed light into the mechanisms by which some nutrients regulate biofilm formation and host colonization.

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Mannitol and the Mannitol-Specific Enzyme IIB Subunit Activate Vibrio cholerae Biofilm Formation

Applied and Environmental Microbiology ◽

10.1128/aem.01184-13 ◽

2013 ◽

Vol 79 (15) ◽

pp. 4675-4683 ◽

Cited By ~ 29

Author(s):

Patrick Ymele-Leki ◽

Laetitia Houot ◽

Paula I. Watnick

Keyword(s):

Vibrio Cholerae ◽

Biofilm Formation ◽

Diarrheal Disease ◽

Ectopic Expression ◽

Phosphotransferase System ◽

Content Type ◽

Regulatory Pathways ◽

B Subunit ◽

Multicomponent Signal ◽

Enzyme I

ABSTRACTVibrio choleraeis a halophilic, Gram-negative rod found in marine environments. Strains that produce cholera toxin cause the diarrheal disease cholera.V. choleraeuse a highly conserved, multicomponent signal transduction cascade known as the phosphoenolpyruvate phosphotransferase system (PTS) to regulate carbohydrate uptake and biofilm formation. Regulation of biofilm formation by the PTS is complex, involving many different regulatory pathways that incorporate distinct PTS components. The PTS consists of the general components enzyme I (EI) and histidine protein (HPr) and carbohydrate-specific enzymes II. Mannitol transport byV. choleraerequires the mannitol-specific EII (EIIMtl), which is expressed only in the presence of mannitol. Here we show that mannitol activatesV. choleraebiofilm formation and transcription of thevpsbiofilm matrix exopolysaccharide synthesis genes. This regulation is dependent on mannitol transport. However, we show that, in the absence of mannitol, ectopic expression of the B subunit of EIIMtlis sufficient to activate biofilm accumulation. Mannitol, a common compatible solute and osmoprotectant of marine organisms, is a main photosynthetic product of many algae and is secreted by algal mats. We propose that the ability ofV. choleraeto respond to environmental mannitol by forming a biofilm may play an important role in habitat selection.

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PafR, a Novel Transcription Regulator, Is Important for Pathogenesis in Uropathogenic Escherichia coli

Infection and Immunity ◽

10.1128/iai.00086-14 ◽

2014 ◽

Vol 82 (10) ◽

pp. 4241-4252 ◽

Cited By ~ 6

Author(s):

Mordechai Baum ◽

Mobarak Watad ◽

Sara N. Smith ◽

Christopher J. Alteri ◽

Noa Gordon ◽

...

Keyword(s):

Escherichia Coli ◽

Biofilm Formation ◽

Gene Knockout ◽

Sensory System ◽

Phosphotransferase System ◽

Knockout Mutant ◽

Double Knockout ◽

Content Type

ABSTRACTThemetVgenomic island in the chromosome of uropathogenicEscherichia coli(UPEC) encodes a putative transcription factor and a sugar permease of the phosphotransferase system (PTS), which are predicted to compose a Bgl-like sensory system. The presence of these two genes, hereby termedpafRandpafP, respectively, has been previously shown to correlate with isolates causing clinical syndromes. We show here that deletion of both genes impairs the ability of the resulting mutant to infect the CBA/J mouse model of ascending urinary tract infection compared to that of the parent strain, CFT073. Expressing the two genes intransin the two-gene knockout mutant complemented full virulence. Deletion of either gene individually generated the same phenotype as the double knockout, indicating that bothpafRandpafPare important to pathogenesis. We screened numerous environmental conditions but failed to detect expression from the promoter that precedes thepafgenesin vitro, suggesting that they arein vivoinduced (ivi). Although PafR is shown here to be capable of functioning as a transcriptional antiterminator, its targets in the UPEC genome are not known. Using microarray analysis, we have shown that expression of PafR from a heterologous promoter in CFT073 affects expression of genes related to bacterial virulence, biofilm formation, and metabolism. Expression of PafR also inhibits biofilm formation and motility. Taken together, our results suggest that thepafgenes are implicated in pathogenesis and that PafR controls virulence genes, in particular biofilm formation genes.

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The Transcription Factor Mlc Promotes Vibrio cholerae Biofilm Formation through Repression of Phosphotransferase System Components

Journal of Bacteriology ◽

10.1128/jb.01639-14 ◽

2014 ◽

Vol 196 (13) ◽

pp. 2423-2430 ◽

Cited By ~ 8

Author(s):

B. S. Pickering ◽

J. E. Lopilato ◽

D. R. Smith ◽

P. I. Watnick

Keyword(s):

Transcription Factor ◽

Vibrio Cholerae ◽

Biofilm Formation ◽

Phosphotransferase System ◽

System Components

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Vibrio cholerae Phosphoenolpyruvate Phosphotransferase System Control of Carbohydrate Transport, Biofilm Formation, and Colonization of the Germfree Mouse Intestine

Infection and Immunity ◽

10.1128/iai.01356-09 ◽

2010 ◽

Vol 78 (4) ◽

pp. 1482-1494 ◽

Cited By ~ 46

Author(s):

Laetitia Houot ◽

Sarah Chang ◽

Cedric Absalon ◽

Paula I. Watnick

Keyword(s):

Vibrio Cholerae ◽

Biofilm Formation ◽

System Control ◽

Cellular Functions ◽

Phosphotransferase System ◽

Germfree Mouse ◽

Abiotic Surfaces ◽

Carbohydrate Transport ◽

Mouse Intestine

ABSTRACT The bacterial phosphoenolpyruvate phosphotransferase system (PTS) is a highly conserved phosphotransfer cascade whose components modulate many cellular functions in response to carbohydrate availability. Here, we further elucidate PTS control of Vibrio cholerae carbohydrate transport and activation of biofilm formation on abiotic surfaces. We then define the role of the PTS in V. cholerae colonization of the adult germfree mouse intestine. We report that V. cholerae colonizes both the small and large intestines of the mouse in a distribution that does not change over the course of a month-long experiment. Because V. cholerae possesses many PTS-independent carbohydrate transporters, the PTS is not essential for bacterial growth in vitro. However, we find that the PTS is essential for colonization of the germfree adult mouse intestine and that this requirement is independent of PTS regulation of biofilm formation. Therefore, competition for PTS substrates may be a dominant force in the success of V. cholerae as an intestinal pathogen. Because the PTS plays a role in colonization of environmental surfaces and the mammalian intestine, we propose that it may be essential to successful transit of V. cholerae through its life cycle of pathogenesis and environmental persistence.

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Contributions of Escherichia coli and its motility to the formation of dual-species biofilms with Vibrio cholerae

Applied and Environmental Microbiology ◽

10.1128/aem.00938-21 ◽

2021 ◽

Author(s):

Hui Wang ◽

Feiyu Li ◽

Li Xu ◽

Hyuntae Byun ◽

JinMing Fan ◽

...

Keyword(s):

Escherichia Coli ◽

Life Cycle ◽

Vibrio Cholerae ◽

Biofilm Formation ◽

Type I ◽

Flagellar Motility ◽

E Coli ◽

Aggregation Formation

Biofilm formation is important in both the environmental and intestinal phases of the Vibrio cholerae life cycle. Nevertheless, most studies of V. cholerae biofilm formation focus on mono-species cultures, whereas nearly all biofilm communities found in nature consist of a variety of microorganisms. Multi-species biofilms formed between V. cholerae and other bacteria in the environment and the interactions that exist between these species are still poorly understood. In this study, the influence of Escherichia coli on the biofilm formation of V. cholerae was studied in the context of both in vitro coculture and in vivo coinfection. To understand the underlying synergistic mechanisms between these two species and to investigate the role of E. coli in V. cholerae biofilm formation, different pathotypes of E. coli and corresponding deletion mutants lacking genes that influence flagella motility, curli fibers, or type I pili were cocultured with V. cholerae . Our findings demonstrate that the presence of commensal E. coli increases biofilm formation at the air-liquid interface in vitro and the generation of biofilm-like multicellular clumps in the mice feces. Examination of laboratory E. coli flagellar-motility mutants Δ fliC and Δ motA in the dual-species biofilm formation suggests that flagellar motility plays an important role in the synergistic interaction and co-aggregation formation between V. cholerae and E. coli . This study facilitates a better understanding of how V. cholerae resides in harsh environments and colonizes the intestine. IMPORTANCE Biofilms play an important role in the V. cholerae life cycle. Until now, mono-species biofilm formation of V. cholerae has been well studied. However, in nature, bacteria live in complex microbial communities, where biofilm is mostly composed of multiple microbial species that interact to cooperate with or compete against each other. Uncovering how V. cholerae forms multi-species biofilm is critical for furthering our understanding of how V. cholerae survives in the environment and transitions to infecting the human host. In this work, the dual-species biofilm between V. cholerae and E. coli was investigated. We demonstrate that the presence of commensal E. coli increased overall biofilm formation. Furthermore, we demonstrate that the motility of E. coli flagella is important for V. cholerae and E. coli to form co-aggregation clumps in dual-species biofilm. These results shed light on a new mechanism for understanding the survival and pathogenesis of V. cholerae .

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The Phosphoenolpyruvate Phosphotransferase System Regulates Vibrio cholerae Biofilm Formation through Multiple Independent Pathways

Journal of Bacteriology ◽

10.1128/jb.00213-10 ◽

2010 ◽

Vol 192 (12) ◽

pp. 3055-3067 ◽

Cited By ~ 60

Author(s):

Laetitia Houot ◽

Sarah Chang ◽

Bradley S. Pickering ◽

Cedric Absalon ◽

Paula I. Watnick

Keyword(s):

Vibrio Cholerae ◽

Biofilm Formation ◽

Minimal Medium ◽

Cellular Functions ◽

Phosphotransferase System ◽

Bacterial Genomes ◽

Regulatory Pathways ◽

Regulatory Circuits ◽

Genes Encoding ◽

Σ Factor

ABSTRACT The bacterial phosphoenolpyruvate phosphotransferase system (PTS) is a highly conserved phosphotransfer cascade that participates in the transport and phosphorylation of selected carbohydrates and modulates many cellular functions in response to carbohydrate availability. It plays a role in the virulence of many bacterial pathogens. Components of the carbohydrate-specific PTS include the general cytoplasmic components enzyme I (EI) and histidine protein (HPr), the sugar-specific cytoplasmic components enzymes IIA (EIIA) and IIB (EIIB), and the sugar-specific membrane-associated multisubunit components enzymes IIC (EIIC) and IID (EIID). Many bacterial genomes also encode a parallel PTS pathway that includes the EI homolog EINtr, the HPr homolog NPr, and the EIIA homolog EIIANtr. This pathway is thought to be nitrogen specific because of the proximity of the genes encoding this pathway to the genes encoding the nitrogen-specific σ factor σ54. We previously reported that phosphorylation of HPr and FPr by EI represses Vibrio cholerae biofilm formation in minimal medium supplemented with glucose or pyruvate. Here we report two additional PTS-based biofilm regulatory pathways that are active in LB broth but not in minimal medium. These pathways involve the glucose-specific enzyme EIIA (EIIAGlc) and two nitrogen-specific EIIA homologs, EIIANtr1 and EIIANtr2. The presence of multiple, independent biofilm regulatory circuits in the PTS supports the hypothesis that the PTS and PTS-dependent substrates have a central role in sensing environments suitable for a surface-associated existence.

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