Quorum Sensing-Dependent Biofilms Enhance Colonization in Vibrio cholerae

Zhu, Jun; Mekalanos, John J.

doi:10.1016/s1534-5807(03)00295-8

Cited by 466 publications

(520 citation statements)

References 43 publications

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“…Genes specifically expressed during biofilm formation in response to environmental conditions or specific mutations have been identified (Whiteley et al, 2001;Schembri et al, 2003;Stanley et al, 2003;Zhu and Mekalanos, 2003), but these studies on whole biofilms lacked the spatial resolution that may be necessary to account for the heterogeneity in metabolic states that can be expected in biofilms (Stewart and Franklin, 2008). Introducing reporters that express unstable green fluorescent proteins can provide insight into the expression of individual genes throughout biofilms (Sternberg et al, 1999;Teal et al, 2006;Stewart and Franklin, 2008).…”

Section: Discussionmentioning

confidence: 99%

Microtoming coupled to microarray analysis to evaluate the spatial metabolic status of Geobacter sulfurreducens biofilms

et al. 2009

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Further insight into the metabolic status of cells within anode biofilms is essential for understanding the functioning of microbial fuel cells and developing strategies to optimize their power output. Cells throughout anode biofilms of Geobacter sulfurreducens reduced the metabolic stains: 5-cyano-2,3-ditolyl tetrazolium chloride and Redox Green, suggesting metabolic activity throughout the biofilm. To compare the metabolic status of cells growing close to the anode versus cells in the outer portion of the anode biofilm, anode biofilms were encased in resin and sectioned into inner (0-20 lm from anode surface) and outer (30-60 lm) fractions. Transcriptional analysis revealed that, at a twofold threshold, 146 genes had significant (Po0.05) differences in transcript abundance between the inner and outer biofilm sections. Only 1 gene, GSU0093, a hypothetical ATP-binding cassette transporter, had significantly higher transcript abundances in the outer biofilm. Genes with lower transcript abundance in the outer biofilm included genes for ribosomal proteins and NADH dehydrogenase, suggesting lower metabolic rates. However, differences in transcript abundance were relatively low (othreefold) and the expression of genes for the tricarboxylic acid cycle enzymes was not significantly lower. Lower expression of genes involved in stress responses in the outer biofilm may reflect the development of low pH near the surface of the anode. The results of this study suggest that cells throughout the biofilm are metabolically active and can potentially contribute to current production. The microtoming/microarray strategy described here may be useful for evaluating gene expression with depth in a diversity of microbial biofilms.

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Section: Discussionmentioning

confidence: 99%

Microtoming coupled to microarray analysis to evaluate the spatial metabolic status of Geobacter sulfurreducens biofilms

et al. 2009

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“…In electron micrographs, VPS appears amorphous, filling the spaces between cells Yildiz and Schoolnik, 1999). Strains overproducing VPS form rugose colonies, exhibit enhanced biofilm formation, and are more resistant to environmental insult Wai et al, 1998;1999;Yildiz and Schoolnik, 1999;Hammer and Bassler, 2003;Zhu and Mekalanos, 2003). In the absence of VPS, V. cholerae produces biofilms under particular conditions (e.g.…”

Section: Discussionmentioning

confidence: 99%

“…Recent important work revealed that OpaR and its V. harveyi homologue LuxR vh also regulate type three secretion and virulence gene expression (Henke and Bassler, 2004). HapR, the V. cholerae OpaR homologue, has also been shown to affect regulation of VPS and other virulence genes (Jobling and Holmes, 1997;Kovacikova and Skorupski, 2002;Hammer and Bassler, 2003;Zhu and Mekalanos, 2003). Like V. parahaemolyticus, spontaneous mutations arise in hapR to promote phase shifting (Zhu et al, 2002;Hammer and Bassler, 2003).…”

Section: Discussionmentioning

confidence: 99%

Genetic determinants of biofilm development of opaque and translucent Vibrio parahaemolyticus

Enos-Berlage

Güvener

Keenan

et al. 2005

Molecular Microbiology

207

193

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SummaryVibrio parahaemolyticus isolates display variation in colony morphology, alternating between opaque (OP) and translucent (TR) cell types. Phase variation is the consequence of genetic alterations in the locus encoding the quorum sensing output regulator OpaR. Here, we show that both cell types form stable, but distinguishable biofilms that differ with respect to attachment and detachment profiles to polystyrene, pellicle formation and stability at the air/medium interface, and submerged biofilm architecture and dispersion at a solid/liquid interface. The pellicle, which is a cohesive mat of cells, was exploited to identify mutants having altered or defective biofilm formation. Transposon insertion mutants were obtained with defects in genes affecting multiple cell surface characteristics, including extracellular polysaccharide, mannose-sensitive haemagglutinin type 4 pili and polar (but not lateral) flagella. Other insertions disrupted genes coding for potential secreted proteins or transporters of secreted proteins, specifically haemolysin co-regulated protein and an RTX toxin-like membrane fusion transporter, as well as potential modifiers of cell surface molecules ( nagAC operon). The pellicle screen also identified mutants with lesions in regulatory genes encoding H-NS, a CsgD-like repressor and an AraC-like protein. This work initiates the characterization of V. parahaemolyticus biofilm formation in the OP and TR cell types and identifies a diverse repertoire of cell surface elements that participate in determining multicellular architecture.

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“…Regulation of quorum sensing by cAMP-CRP expressed as the A 570 /OD 600 ratio (Zhu & Mekalanos, 2003). Strains were grown in 5 ml LB medium for 16 h, diluted 1 : 50 in fresh medium and transferred to 96-well flat-bottom microtitre plates.…”

Section: Methodsmentioning

confidence: 99%

“…The HapR regulator plays a pivotal role in regulating virulence gene expression and biofilm development (Hammer & Bassler, 2003;Zhu & Mekalanos, 2003). At low cell density the regulator AphA activates expression of TcpP/H (Häse & Kovacikova & Skorupski, 2001), which in concert with a second pair of transmembrane regulators (ToxR/S) activates expression of the soluble regulator ToxT (Miller et al, 1989).…”

Section: Introductionmentioning

confidence: 99%

The cyclic AMP receptor protein modulates quorum sensing, motility and multiple genes that affect intestinal colonization in Vibrio cholerae

et al. 2007

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Vibrio cholerae is the causative agent of cholera, which continues to be a major public health concern in Asia, Africa and Latin America. The bacterium can persist outside the human host and alternates between planktonic and biofilm community lifestyles. Transition between the different lifestyles is mediated by multiple signal transduction pathways including quorum sensing. Expression of the Zn-metalloprotease haemagglutinin (HA)/protease is subject to a dual regulation which involves the quorum-sensing regulator HapR and the cAMP receptor protein. In a previous study, we observed that a mutant defective in the cAMP-receptor protein (CRP) expressed lower levels of HapR. To further investigate the role of CRP in modulating HapR and other signal transduction pathways, we performed global gene expression profiling of a Dcrp mutant of El Tor biotype V. cholerae. Here we show that CRP is required for the biosynthesis of cholera autoinducer 1 (CAI-1) and affects the expression of multiple HapR-regulated genes. As expected, the Dcrp mutant produced more cholera toxin and enhanced biofilm. Expression of flagellar genes, reported to be affected in DhapR mutants, was diminished in the Dcrp mutant. However, an epistasis analysis indicated that cAMP-CRP affects motility by a mechanism independent of HapR. Inactivation of crp inhibited the expression of multiple genes reported to be strongly induced in vivo and to affect the ability of V. cholerae to colonize the small intestine and cause disease. These genes included ompU, ompT and ompW encoding outer-membrane proteins, the alternative sigma factor s E required for intestinal colonization, and genes involved in anaerobic energy metabolism. Our results indicate that CRP plays a crucial role in the V. cholerae life cycle by affecting quorum sensing and multiple genes required for survival of V. cholerae in the human host and the environment.

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Quorum Sensing-Dependent Biofilms Enhance Colonization in Vibrio cholerae

Cited by 466 publications

References 43 publications

Microtoming coupled to microarray analysis to evaluate the spatial metabolic status of Geobacter sulfurreducens biofilms

Microtoming coupled to microarray analysis to evaluate the spatial metabolic status of Geobacter sulfurreducens biofilms

Genetic determinants of biofilm development of opaque and translucent Vibrio parahaemolyticus

The cyclic AMP receptor protein modulates quorum sensing, motility and multiple genes that affect intestinal colonization in Vibrio cholerae

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