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Vanessa Sperandio

Publications and source records attributed to Vanessa Sperandio.

At least 19 recordsLinked to original sources
PubMed2007

A novel two-component signaling system that activates transcription of an enterohemorrhagic Escherichia coli effector involved in remodeling of host actin.

Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is responsible for worldwide outbreaks of bloody diarrhea, hemorrhagic colitis, and life-threatening hemolytic uremic syndrome. After colonizing the large intestine, EHEC forms attaching and effacing (AE) lesions on intestinal epithelial cells. These lesions cause destruction of the microvilli and elicit actin rearrangement to form pedestals that cup each bacterium individually. EHEC responds to a signal produced by the intestinal microbial flora, autoinducer-3 (AI-3), and the host hormones epinephrine and norepinephrine to activate transcription of the genes involved in AE lesion formation. These three signals, involved in interkingdom communication, are sensed by bacterial sensor kinases. Here we describe a novel two-component system, QseEF (quorum-sensing E. coli regulators E and F), which is part of the AI-3/epinephrine/norepinephrine signaling system. QseE is the sensor kinase and QseF the response regulator. The qseEF genes are cotranscribed, and transcription of qseEF is activated by epinephrine through the QseC sensor. A qseF mutant does not form AE lesions. QseF activates transcription of the gene encoding EspFu, an effector protein translocated to the host cell by the EHEC, which mimics a eukaryotic SH2/SH3 adapter protein to engender actin polymerization during pedestal formation. Expression of the espFu gene from a plasmid restored AE lesion formation to the qseF mutant, suggesting that lack of espFu expression in this mutant was responsible for the loss of pedestal formation. These findings suggest the QseEF is a two-component system involved in the regulation of AE lesion formation by EHEC.

Actins↗
PubMed2007

Quorum sensing by enteric pathogens.

PURPOSE OF REVIEW: This review presents advances in our understanding of how pathogenic, enteric bacteria use quorum sensing to regulate several traits that allow them to establish and maintain infection in their host, including motility, biofilm formation, and virulence-specific genes. RECENT FINDINGS: Quorum sensing in enteric bacteria has been elusive for a long time. Recent data indicate that enteric bacteria use several quorum-sensing mechanisms including the LuxR-I quorum-sensing system, the LuxS/AI-2 system, and the AI-3/epinephrine/norepinephrine system to assess their environment and to recognize the host environment. These systems allow bacteria to communicate across species boundaries, and the AI-3/epinephrine/norepinephrine system is involved in interkingdom signaling. SUMMARY: Recent developments in our understanding of the molecular and biochemical mechanisms involved in quorum sensing as well as the chemical signal(s) to which bacteria respond and their corresponding physiological responses will improve our understanding of bacterial pathogenesis and microbial flora-host interactions, and potentially lead to novel strategies for combating infection.

Animals↗
PubMed2006

The QseC sensor kinase: a bacterial adrenergic receptor.

Quorum sensing is a cell-to-cell signaling mechanism in which bacteria respond to hormone-like molecules called autoinducers (AIs). The AI-3 quorum-sensing system is also involved in interkingdom signaling with the eukaryotic hormones epinephrine/norepinephrine. This signaling activates transcription of virulence genes in enterohemorrhagic Escherichia coli O157:H7. However, this signaling system has never been shown to be involved in virulence in vivo, and the bacterial receptor for these signals had not been identified. Here, we show that the QseC sensor kinase is a bacterial receptor for the host epinephrine/norepinephrine and the AI-3 produced by the gastrointestinal microbial flora. We also found that an alpha-adrenergic antagonist can specifically block the QseC response to these signals. Furthermore, we demonstrated that a qseC mutant is attenuated for virulence in a rabbit animal model, underscoring the importance of this signaling system in virulence in vivo. Finally, an in silico search found that the periplasmic sensing domain of QseC is conserved among several bacterial species. Thus, QseC is a bacterial adrenergic receptor that activates virulence genes in response to interkingdom cross-signaling. We anticipate that these studies will be a starting point in understanding bacterial-host hormone signaling at the biochemical level. Given the role that this system plays in bacterial virulence, further characterization of this unique signaling mechanism may be important for developing novel classes of antimicrobials.

Animals↗
PubMed2006

Quorum sensing in Escherichia coli and Salmonella.

Quorum sensing in Escherichia coli and Salmonella has been an elusive topic for a long time. However, in the past 8 years, several research groups have demonstrated that these bacteria use several quorum-sensing systems, such as: the luxS/AI-2, AI-3/epinephrine/norepinephrine, indole, and the LuxR homolog SdiA to achieve intercellular signaling. The majority of these signaling systems are involved in interspecies communication, and the AI-3/epinephrine/norepinephrine signaling system is also involved in interkingdom communication. Both E. coli and Salmonella reside in the human intestine, which is the largest and most complex environment in the mammalian host. The observation that these bacteria evolved quorum-sensing systems primarily involved in interspecies communication may constitute an adaptation to this environment. The gastrointestinal tract harbors a high density and diversity of bacterial cells, with the majority of the flora residing in the colon (10(11)-10(12) bacterial cells/ml). Given the enormous number and diversity of bacteria inhabiting the gastrointestinal environment, it should not be surprising that the members of this community communicate amongst themselves and with the host itself to coordinate a variety of adaptive processes.

Bacterial Proteins↗
PubMed2006

Quorum sensing: the many languages of bacteria.

In the conventional view of prokaryotic existence, bacteria live unicellularly, with responses to external stimuli limited to the detection of chemical and physical signals of environmental origin. This view of bacteriology is now recognized to be overly simplistic, because bacteria communicate with each other through small 'hormone-like' organic compounds referred to as autoinducers. These bacterial cell-to-cell signaling systems were initially described as mechanisms through which bacteria regulate gene expression via cell density and, therefore, they have been collectively termed quorum sensing. The functions controlled by quorum sensing are varied and reflect the needs of a particular species of bacteria to inhabit a given niche. Three major quorum-sensing circuits have been described: one used primarily by Gram-negative bacteria, one used primarily by Gram-positive bacteria, and one that has been proposed to be universal.

4-Butyrolactone↗
PubMed2006

Autoinducer 3 and epinephrine signaling in the kinetics of locus of enterocyte effacement gene expression in enterohemorrhagic Escherichia coli.

Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is responsible for causing outbreaks of bloody diarrhea and hemolytic-uremic syndrome throughout the world. The locus of enterocyte effacement (LEE) consists of five major operons and is required for the formation of attaching and effacing lesions that disrupt intestinal epithelial microvilli. We have previously reported that expression of EHEC LEE genes is regulated by the luxS quorum-sensing system. The luxS gene in EHEC affects the production of autoinducer 3 (AI-3), which activates the LEE. Epinephrine and norepinephrine also activate the LEE in a manner similar to that of AI-3. Previous studies of quorum-sensing regulation of LEE transcription have thus far been restricted to using reporter systems in an E. coli K-12 background. Here, we examined the kinetics of LEE gene transcription, protein expression, and function of the LEE type III secretion apparatus in wild-type (WT) EHEC and an isogenic luxS mutant. The results revealed that the luxS mutant had diminished transcription from the LEE promoters during the mid-exponential growth phase; decreased protein levels of EscJ, Tir, and EspA; and reduced secretion of EspA and EspB. The luxS mutation also caused a delay in the formation of attaching and effacing lesions on cultured epithelial cells compared to the wild type. Epinephrine enhanced LEE expression in both the WT and the luxS mutant, but the WT still exhibited greater LEE activation. The results suggest a possible synergistic relationship between AI-3 and epinephrine. The combined effects of these two signaling molecules may lead to greater LEE expression and a more efficient infection.

Bacterial Proteins↗
PubMed2006

Bundle-forming pili and EspA are involved in biofilm formation by enteropathogenic Escherichia coli.

Microcolony formation is one of the initial steps in biofilm development, and in enteropathogenic Escherichia coli (EPEC) it is mediated by several adhesins, including the bundle-forming pilus (BFP) and the EspA filament. Here we report that EPEC forms biofilms on plastic under static conditions and a flowthrough continuous culture system. The abilities of several EPEC isogenic mutants to form biofilms were assessed. Adhesins such as BFP and EspA, important in microcolony formation on epithelial cells, are also involved in bacterial aggregation during biofilm formation on abiotic surfaces. Mutants that do not express BFP or EspA form more-diffuse biofilms than does the wild type. We also determined, using gfp transcriptional fusions, that, consistent with the role of these adhesins in biofilms, the genes encoding BFP and EspA are expressed during biofilm formation. Finally, expression of espA is controlled by a quorum-sensing (QS) regulatory mechanism, and the EPEC qseA QS mutant also forms altered biofilms, suggesting that this signaling mechanism plays an important role in EPEC biofilm development. Taken together, these studies allowed us to propose a model of EPEC biofilm formation.

Biofilms↗
PubMed2006

AI-3 synthesis is not dependent on luxS in Escherichia coli.

The quorum-sensing (QS) signal autoinducer-2 (AI-2) has been proposed to promote interspecies signaling in a broad range of bacterial species. AI-2 is spontaneously derived from 4,5-dihydroxy-2,3-pentanedione that, along with homocysteine, is produced by cleavage of S-adenosylhomocysteine (SAH) and S-ribosylhomocysteine by the Pfs and LuxS enzymes. Numerous phenotypes have been attributed to AI-2 QS signaling using luxS mutants. We have previously reported that the luxS mutation also affects the synthesis of the AI-3 autoinducer that activates enterohemorrhagic Escherichia coli virulence genes. Here we show that several species of bacteria synthesize AI-3, suggesting a possible role in interspecies bacterial communication. The luxS mutation leaves the cell with only one pathway, involving oxaloacetate and l-glutamate, for de novo synthesis of homocysteine. The exclusive use of this pathway for homocysteine production appears to alter metabolism in the luxS mutant, leading to decreased levels of AI-3. The addition of aspartate and expression of an aromatic amino acid transporter, as well as a tyrosine-specific transporter, restored AI-3-dependent phenotypes in an luxS mutant. The defect in AI-3 production, but not in AI-2 production, in the luxS mutant was restored by expressing the Pseudomonas aeruginosa S-adenosylhomocysteine hydrolase that synthesizes homocysteine directly from SAH. Furthermore, phenotype microarrays revealed that the luxS mutation caused numerous metabolic deficiencies, while AI-3 signaling had little effect on metabolism. This study examines how AI-3 production is affected by the luxS mutation and explores the roles of the LuxS/AI-2 system in metabolism and QS.

Amino Acid Transport Systems↗
PubMed2005

Transcriptional regulation of flhDC by QseBC and sigma (FliA) in enterohaemorrhagic Escherichia coli.

Enterohaemorrhagic Escherichia coli (EHEC) serotype O157:H7, the causative agent of haemorrhagic colitis, has been shown to utilize a cell-to-cell signalling system to regulate gene expression. We have previously reported that the quorum sensing E. coli regulators B and C (QseBC) may act as a two-component system in EHEC to transcriptionally regulate the expression of flagella and motility through flhDC, the master regulator of flagella and motility genes. Here, we performed deletion analyses using the flhDC promoter in order to determine the minimal promoter regions necessary for QseBC transcriptional activation. We also performed electrophoretic mobility shift assays, competition experiments and DNaseI footprints, which suggest that QseB directly binds the flhDC promoter at high- and low-affinity binding sites. These analyses have allowed us to determine the potential consensus sequence to which QseB binds in order to regulate transcription. Additionally, we mapped the transcriptional start site of flhDC responsive to QseBC, leading to the identification of a conserved FliA (sigma28) consensus sequence. These results suggest that FliA (sigma28), a class 2 flagellar gene, may be aiding in the transcriptional initiation of class 1 genes (flhDC) in EHEC. In order to further characterize the role of FliA (sigma28) in transcription of the flhDC promoter, we constructed a fliA isogenic mutant in EHEC. The flhDC::lacZ transcriptional fusion showed decreased activity in the fliA mutant compared with wild-type and complemented strains. Taken together, these results indicate that transcriptional initiation at the flhDC promoter by QseBC appears to be complex and dependent on the presence of FliA (sigma28).

Bacterial Proteins↗
PubMed2005

Transcriptional autoregulation by quorum sensing Escherichia coli regulators B and C (QseBC) in enterohaemorrhagic E. coli (EHEC).

The cell-to-cell communication system referred to as quorum sensing (QS) is based on the principle that bacteria secrete hormone-like compounds referred to as autoinducers. Upon reaching a threshold concentration, these autoinducers interact with transcription factors to regulate gene expression. We previously reported that enterohaemorrhagic Escherichia coli (EHEC), which is responsible for outbreaks of bloody diarrhoea, utilizes a QS system to regulate gene transcription. We have also previously shown that the quorum sensing E. coli regulators B and C (QseBC) may act as a two-component system to transcriptionally regulate the expression of flagella and motility. Here, using a reverse transcription polymerase chain reaction (RT-PCR), we show that qseBC are transcribed in an operon. Furthermore, using a qseBC::lacZ transcriptional fusion, we observed that QseB autoactivates its own transcription. In addition, the transcriptional start site of the qseBC promoter responsive to QseBC was mapped, and single-copy and multicopy deletion analyses were performed to determine the minimal region necessary for QseB transcriptional activation. These data allowed us to map an additional transcriptional start site for the qseBC promoter which may allow for a basal level of QseBC expression. Finally, electrophoretic mobility shift assays, competition experiments and DNase I footprints were performed and demonstrated that QseB directly binds to two sites in its own promoter. These results indicate that QseB may act to autoregulate its own transcription through binding to low- and high-affinity sites found in its promoter.

Escherichia coli O157↗
PubMed2005

Colonization of gnotobiotic piglets by a luxS mutant strain of Escherichia coli O157:H7.

Gnotobiotic piglets inoculated with Escherichia coli O157:H7, its luxS mutant derivative, or nonpathogenic E. coli were evaluated for attaching and effacing lesions. Although no differences in clinical symptoms were seen between pigs inoculated with the parent and those inoculated with the luxS mutant, the luxS mutant-inoculated pigs had a lower frequency of attaching and effacing lesions in the spiral colon than parent strain-inoculated pigs.

Animals↗
PubMed2005

Events at the host-microbial interface of the gastrointestinal tract III. Cell-to-cell signaling among microbial flora, host, and pathogens: there is a whole lot of talking going on.

Humans have an important association with their intestinal microbial flora. The microbial flora helps to shape the mammalian innate immune system, absorbs nutrients, and plays an intricate role on intestinal development. Microbes and mammals communicate with each other through an array of hormone and hormonelike chemical compounds. These "signals," however, are hijacked by bacterial pathogens, such as enterohemorrhagic Eschrichia coli (EHEC), to activate its virulence genes, colonize the host, and start the disease process. This review explores the cell-to-cell signaling events in the gastrointestinal tract that lead EHEC to regulate its virulence genes in a coordinate manner.

Bacteria↗
PubMed2004

Striking a balance: inter-kingdom cell-to-cell signaling, friendship or war?

There is a growing body of evidence suggesting that bacterial signaling molecules are involved in cross-kingdom communication with eukaryotic cells. Bacterial signaling molecules can enter eukaryotic cells and appear to modulate immune responses to bacterial pathogens. Recently, an enticing study demonstrated that mammalian airway epithelial cells have the ability to inactivate these bacterial signaling molecules, suggesting that this activity might have a role in the host innate response against bacterial infections.

Animals↗
PubMed2004

Modulation of enteropathogenic Escherichia coli virulence by quorum sensing.

Enteropathogenic Escherichia coli (EPEC) produces a lesion on epithelial cells called the attaching and effacing (AE) lesion. All genes necessary for AE are encoded within the locus of enterocyte effacement (LEE). EPEC also adheres in a characteristic pattern to epithelial cells by forming microcolonies, usually referred to as localized adherence (LA). LA is mediated by the bundle-forming pilus and flagella. The LEE genes are directly activated by the LEE-encoded regulator (Ler). Transcription of Ler is under the control of Per, integration host factor, Fis, BipA, and quorum sensing (QS), specifically through the luxS system. QS activates expression of the LEE genes in EPEC, with QseA activating transcription of ler. Here we report that transcription of the LEE genes and type III secretion are diminished in both luxS and qseA mutants. Transcription of the LEE genes is affected in both mutants mostly during the mid-exponential phase of growth. Transcription of qseA itself is diminished throughout growth in a luxS mutant and is under autorepression. Furthermore, QS activation of type III secretion is independent of per, given that QseA still activates type III secretion in a per mutant strain. Both mutants are deficient in adherence to epithelial cells and form smaller microcolonies. Several factors may contribute to this abnormal behavior: transcription of LEE genes and type III secretion are diminished, and expression of flagella and Per is altered in both mutants. These results suggest that QS is involved in modulating the regulation of the EPEC virulence genes.

Bacterial Adhesion↗
PubMed2004

Cell-to-cell signaling in intestinal pathogens.

In the conventional view of prokaryotic life, bacteria live a unicellular existence, with responses to external stimuli limited to the detection of chemical and physical signals of environmental origin. This view of bacteriology is now recognized as overly simplistic, because bacteria communicate with each other through small "hormone-like" organic compounds referred to as autoinducers (Als). These bacterial cell-to-cell signaling systems were initially described as mechanisms through which bacteria regulate gene expression via cell density, and, therefore, they have been named quorum sensing. When the Als reach a threshold concentration, they interact with regulatory proteins, thereby driving bacterial gene expression. Bacterial intercellular communication provides a mechanism for the regulation of gene expression resulting in coordinated population behavior. The functions controlled by quorum sensing are varied and reflect the needs of a particular species of bacteria inhabiting a given niche. Quorum sensing-controlled processes include bioluminescence, virulence factor expression, biofilm development, and conjugation among others. Enteric pathogens use quorum sensing to regulate genes involved in virulence, such as motility, and type III secretion. Quorum sensing is utilized to sense the presence of the normal intestinal flora and to warrant successful colonization of the host.

Bacterial Proteins↗
PubMed2003

Bacteria-host communication: the language of hormones.

The interbacterial communication system known as quorum sensing (QS) utilizes hormone-like compounds referred to as autoinducers to regulate bacterial gene expression. Enterohemorrhagic Escherichia coli (EHEC) serotype O157:H7 is the agent responsible for outbreaks of bloody diarrhea in several countries. We previously proposed that EHEC uses a QS regulatory system to "sense" that it is within the intestine and activate genes essential for intestinal colonization. The QS system used by EHEC is the LuxS/autoinducer 2 (AI-2) system extensively involved in interspecies communication. The autoinducer AI-2 is a furanosyl borate diester whose synthesis depends on the enzyme LuxS. Here we show that an EHEC luxS mutant, unable to produce the bacterial autoinducer, still responds to a eukaryotic cell signal to activate expression of its virulence genes. We have identified this signal as the hormone epinephrine and show that beta- and alpha-adrenergic antagonists can block the bacterial response to this hormone. Furthermore, using purified and in vitro synthesized AI-2 we showed that AI-2 is not the autoinducer involved in the bacterial signaling. EHEC produces another, previously undescribed autoinducer (AI-3) whose synthesis depends on the presence of LuxS. These results imply a potential cross-communication between the luxS/AI-3 bacterial QS system and the epinephrine host signaling system. Given that eukaryotic cell-to-cell signaling typically occurs through hormones, and that bacterial cell-to-cell signaling occurs through QS, we speculate that QS might be a "language" by which bacteria and host cells communicate.

Bacterial Proteins↗