scholarly journals The Extracellular Nuclease Dns and Its Role in Natural Transformation of Vibrio cholerae

2008 ◽  
Vol 190 (21) ◽  
pp. 7232-7240 ◽  
Author(s):  
Melanie Blokesch ◽  
Gary K. Schoolnik

ABSTRACT Free extracellular DNA is abundant in many aquatic environments. While much of this DNA will be degraded by nucleases secreted by the surrounding microbial community, some is available as transforming material that can be taken up by naturally competent bacteria. One such species is Vibrio cholerae, an autochthonous member of estuarine, riverine, and marine habitats and the causative agent of cholera, whose competence program is induced after colonization of chitin surfaces. In this study, we investigate how Vibrio cholerae's two extracellular nucleases, Xds and Dns, influence its natural transformability. We show that in the absence of Dns, transformation frequencies are significantly higher than in its presence. During growth on a chitin surface, an increase in transformation efficiency was found to correspond in time with increasing cell density and the repression of dns expression by the quorum-sensing regulator HapR. In contrast, at low cell density, the absence of HapR relieves dns repression, leading to the degradation of free DNA and to the abrogation of the transformation phenotype. Thus, as cell density increases, Vibrio cholerae undergoes a switch from nuclease-mediated degradation of extracellular DNA to the uptake of DNA by bacteria induced to a state of competence by chitin. Taken together, these results suggest the following model: nuclease production by low-density populations of V. cholerae might foster rapid growth by providing a source of nucleotides for the repletion of nucleotide pools. In contrast, the termination of nuclease production by static, high-density populations allows the uptake of intact DNA and coincides with a phase of potential genome diversification.

2008 ◽  
Vol 190 (7) ◽  
pp. 2527-2536 ◽  
Author(s):  
Christopher M. Waters ◽  
Wenyun Lu ◽  
Joshua D. Rabinowitz ◽  
Bonnie L. Bassler

ABSTRACT Two chemical signaling systems, quorum sensing (QS) and 3′,5′-cyclic diguanylic acid (c-di-GMP), reciprocally control biofilm formation in Vibrio cholerae. QS is the process by which bacteria communicate via the secretion and detection of autoinducers, and in V. cholerae, QS represses biofilm formation. c-di-GMP is an intracellular second messenger that contains information regarding local environmental conditions, and in V. cholerae, c-di-GMP activates biofilm formation. Here we show that HapR, a major regulator of QS, represses biofilm formation in V. cholerae through two distinct mechanisms. HapR controls the transcription of 14 genes encoding a group of proteins that synthesize and degrade c-di-GMP. The net effect of this transcriptional program is a reduction in cellular c-di-GMP levels at high cell density and, consequently, a decrease in biofilm formation. Increasing the c-di-GMP concentration at high cell density to the level present in the low-cell-density QS state restores biofilm formation, showing that c-di-GMP is epistatic to QS in the control of biofilm formation in V. cholerae. In addition, HapR binds to and directly represses the expression of the biofilm transcriptional activator, vpsT. Together, our results suggest that V. cholerae integrates information about the vicinal bacterial community contained in extracellular QS autoinducers with the intracellular environmental information encoded in c-di-GMP to control biofilm formation.


2009 ◽  
Vol 191 (22) ◽  
pp. 6911-6917 ◽  
Author(s):  
Brooke A. Jude ◽  
Raquel M. Martinez ◽  
Karen Skorupski ◽  
Ronald K. Taylor

ABSTRACT Vibrio cholerae is the etiologic agent of cholera in humans. Intestinal colonization occurs in a stepwise fashion, initiating with attachment to the small intestinal epithelium. This attachment is followed by expression of the toxin-coregulated pilus, microcolony formation, and cholera toxin (CT) production. We have recently characterized a secreted attachment factor, GlcNAc binding protein A (GbpA), which functions in attachment to environmental chitin sources as well as to intestinal substrates. Studies have been initiated to define the regulatory network involved in GbpA induction. At low cell density, GbpA was detected in the culture supernatant of all wild-type (WT) strains examined. In contrast, at high cell density, GbpA was undetectable in strains that produce HapR, the central regulator of the cell density-dependent quorum-sensing system of V. cholerae. HapR represses the expression of genes encoding regulators involved in V. cholerae virulence and activates the expression of genes encoding the secreted proteases HapA and PrtV. We show here that GbpA is degraded by HapA and PrtV in a time-dependent fashion. Consistent with this, ΔhapA ΔprtV strains attach to chitin beads more efficiently than either the WT or a ΔhapA ΔprtV ΔgbpA strain. These results suggest a model in which GbpA levels fluctuate in concert with the bacterial production of proteases in response to quorum-sensing signals. This could provide a mechanism for GbpA-mediated attachment to, and detachment from, surfaces in response to environmental cues.


2021 ◽  
Author(s):  
HA Kittredge ◽  
SE Evans

AbstractThe exchange of genes between potentially unrelated bacteria is termed horizontal gene transfer (HGT) and is a driving force in bacterial evolution. Natural transformation is one mechanism of HGT where extracellular DNA (eDNA) from the environment is recombined into a host genome. The widespread conservation of transformation in bacterial lineages implies there is a fitness benefit. However, the nature of these benefits and the evolutionary origins of transformation are still unknown. Here, I examine how ∼330 generations or 100 days of serial passage in either constant or increasing salinities impacts the growth rate and transformation efficiency of Pseudomonas stutzeri. While the growth rate generally improved in response to serial transfer, the transformation efficiency of the evolved lineages varied extensively, with only 39-64% of populations undergoing transformation at the end of adaptive evolution. In comparison, 100% of the ancestral populations were able to undergo natural transformation. I also found that evolving P. stutzeri with different cell lysates (or populations of dead cells) minimally affected the growth rate and transformation efficiency, especially in comparison to the pervasiveness with which transformation capacity was lost across the evolved populations. Taken together, I show that the efficiency of eDNA uptake changes over relatively rapid timescales, suggesting that transformation is an adaptive and selectable trait that could be lost in environments where it is not beneficial.


2020 ◽  
Vol 10 (3) ◽  
pp. 186-191
Author(s):  
Sara M. Van Bonn ◽  
Sebastian P. Schraven ◽  
Tobias Schuldt ◽  
Markus M. Heimesaat ◽  
Robert Mlynski ◽  
...  

AbstractWe report a case of a chronic mesotympanic otitis media with a smelly purulent secretion from both ears and recurrent otalgia over the last five years in a six-year-old girl after swimming in the German Baltic Sea. Besides Staphylococcus aureus a non-O1/non-O139 Vibrio cholerae strain could be isolated from patient samples. An antibiotic therapy with ciprofloxacin and ceftriaxone was administered followed by atticotomy combined with tympanoplasty. We conclude that V. cholerae should not be overlooked as a differential diagnosis to otitis infections, especially when patients present with extra-intestinal infections after contact with brackish- or saltwater aquatic environments.


Blood ◽  
1997 ◽  
Vol 90 (1) ◽  
pp. 234-243 ◽  
Author(s):  
Lindsey J. White ◽  
Bradford W. Ozanne ◽  
Pierre Graber ◽  
Jean-Pierre Aubry ◽  
Jean-Yves Bonnefoy ◽  
...  

Abstract Human CD23 is a 45-kD type II membrane glycoprotein, which functions as a low-affinity receptor for IgE and as a ligand for the CD21 and CD11b/CD11c differentiation antigens. CD23 is released from the surface of cells as soluble fragments, and a 25-kD species of soluble CD23 (sCD23) appears to act as a multifunctional cytokine. In this report, sCD23 is shown to sustain the growth of low cell density cultures of a human pre-B–acute lymphocytic leukemia cell line, SMS-SB: no other cytokine tested was able to induce this effect. Flow cytometric analysis indicates that sCD23 acts to prevent apoptosis of SMS-SB cells. SMS-SB cells cultured at low cell density possess low levels of bcl-2 protein. Addition of sCD23 to cells at low cell density maintained bcl-2 expression at levels equivalent to those observed in SMS-SB cells cultured at higher cell densities. No CD23 mRNA was found in SMS-SB cells, ruling out an autocrine function for CD23 in this cell line model. Although SMS-SB cells do not express the known receptors for CD23, namely CD21, CD11b-CD18, or CD11c-CD18, the cells specifically bind CD23-containing liposomes, but not glycophorin-containing liposomes. Binding of CD23-containing liposomes is inhibited by anti-CD23 but not by anti-CD21 or anti-CD11b/c monoclonal antibodies. The data show that sCD23 prevents apoptosis of the SMS-SB cell line by acting through a novel receptor.


1992 ◽  
Vol 7 (1) ◽  
pp. 43-46 ◽  
Author(s):  
MASAHIKO NISHIMURA ◽  
KUMIKO KITA-TSUKAMOTO ◽  
KAZUHIRO KOGURE ◽  
KOUICHI OHWADA

2018 ◽  
Vol 5 (5) ◽  
pp. 171792 ◽  
Author(s):  
Jesse P. Harrison ◽  
Carl Boardman ◽  
Kenneth O'Callaghan ◽  
Anne-Marie Delort ◽  
Jim Song

Plastic litter is encountered in aquatic ecosystems across the globe, including polar environments and the deep sea. To mitigate the adverse societal and ecological impacts of this waste, there has been debate on whether ‘biodegradable' materials should be granted exemptions from plastic bag bans and levies. However, great care must be exercised when attempting to define this term, due to the broad and complex range of physical and chemical conditions encountered within natural ecosystems. Here, we review existing international industry standards and regional test methods for evaluating the biodegradability of plastics within aquatic environments (wastewater, unmanaged freshwater and marine habitats). We argue that current standards and test methods are insufficient in their ability to realistically predict the biodegradability of carrier bags in these environments, due to several shortcomings in experimental procedures and a paucity of information in the scientific literature. Moreover, existing biodegradability standards and test methods for aquatic environments do not involve toxicity testing or account for the potentially adverse ecological impacts of carrier bags, plastic additives, polymer degradation products or small (microscopic) plastic particles that can arise via fragmentation. Successfully addressing these knowledge gaps is a key requirement for developing new biodegradability standard(s) for lightweight carrier bags.


2019 ◽  
Author(s):  
Andrew A. Bridges ◽  
Bonnie L. Bassler

AbstractVibrio cholerae possesses multiple quorum-sensing systems that control virulence and biofilm formation among other traits. At low cell densities, when quorum-sensing autoinducers are absent, V. cholerae forms biofilms. At high cell densities, when autoinducers have accumulated, biofilm formation is repressed and dispersal occurs. Here, we focus on the roles of two well-characterized quorum-sensing autoinducers that function in parallel. One autoinducer, called CAI-1, is used to measure vibrio abundance, and the other autoinducer, called AI-2, is a broadly-made universal autoinducer that is presumed to enable V. cholerae to assess the total bacterial cell density of the vicinal community. The two V. cholerae autoinducers funnel information into a shared signal relay pathway. This feature of the quorum-sensing system architecture has made it difficult to understand how specific information can be extracted from each autoinducer, how the autoinducers might drive distinct output behaviors, and in turn, how the bacteria use quorum sensing to distinguish self from other in bacterial communities. We develop a live-cell biofilm formation and dispersal assay that allows examination of the individual and combined roles of the two autoinducers in controlling V. cholerae behavior. We show that the quorum-sensing system works as a coincidence detector in which both autoinducers must be present simultaneously for repression of biofilm formation to occur. Within that context, the CAI-1 quorum-sensing pathway is activated when only a few V. cholerae cells are present, whereas the AI-2 pathway is activated only at much higher cell density. The consequence of this asymmetry is that exogenous sources of AI-2, but not CAI-1, contribute to satisfying the coincidence detector to repress biofilm formation and promote dispersal. We propose that V. cholerae uses CAI-1 to verify that some of its kin are present before committing to the high-cell-density quorum-sensing mode, but it is, in fact, the universal autoinducer AI-2, that sets the pace of the V. cholerae quorum-sensing program. This first report of unique roles for the different V. cholerae autoinducers suggests that detection of self fosters a distinct outcome from detection of other.


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