A Novel Role for Enzyme I of the Vibrio cholerae Phosphoenolpyruvate Phosphotransferase System in Regulation of Growth in a Biofilm

Abstract

Glucose is a universal energy source and a potent inducer of surface colonization for many microbial species. Highly efficient sugar assimilation pathways ensure successful competition for this preferred carbon source. One such pathway is the phosphoenolpyruvate phosphotransferase system (PTS), a multicomponent sugar transport system that phosphorylates the sugar as it enters the cell. Components required for transport of glucose through the PTS include enzyme I, histidine protein, enzyme IIA Glc , and enzyme IIBC Glc . In Escherichia coli, components of the PTS fulfill many regulatory roles, including regulation of nutrient scavenging and catabolism, chemotaxis, glycogen utilization, catabolite repression, and inducer exclusion. We previously observed that genes encoding the components of the Vibrio cholerae PTS were coregulated with the vps genes, which are required for synthesis of the biofilm matrix exopolysaccharide. In this work, we identify the PTS components required for transport of glucose and investigate the role of each of these components in regulation of biofilm formation. Our results establish a novel role for the phosphorylated form of enzyme I in specific regulation of biofilm-associated growth. As the PTS is highly conserved among bacteria, the enzyme I regulatory pathway may be relevant to a number of biofilm-based infections.

Integration of Global Regulatory Mechanisms Controlling Vibrio Cholerae Behavior

“…Biofilm formation is also modulated by interplay between the positive transcription regulators, VpsT (Casper-Lindley & Yildiz, 2004) and VpsR (Yildiz et al, 2001), and the negative regulator, CytR (Haugo & Watnick, 2002). In addition, vps expression is modulated by the PhoBR two-component regulatory system (Pratts et al, 2009(Pratts et al, , 2010Sultan et al, 2010) and by components of the PTS phosphoryl cascade (Houot et al, 2008;, 2010a.…”

“…Pretreatment of mice with commensal bacteria engineered to express CAI-1 affords protection against a cholera challenge (Duan and March, 2010). A second set of attractive targets are the PTS components that modulate biofilm formation (Houot et al, 2008(Houot et al, , 2010(Houot et al, , 2010a. A comparative genomic analysis of 202 fully sequenced genomes (174 bacterial, 19 archaeal, and 9 eukaryotic) did not reveal components of the PTS system in eukaryotic cells (Barabote et al, 2005).…”

“…For instance, in minimal medium, phosphoryl transfer from EI to HPr and FPr represses vps expression (Houot et al, 2008); in LB medium, glucose-specific EIIA glc and nitrogen-specific EIIA Ntr1 and EIIA Ntr2 activate and repress vps expression, respectively (Houot et al, 2010). …”

Antibiotic Resistance, Biofilms and Quorum Sensing in Acinetobacter Species

“…Rather, according to UniProtKB annotations, it is a putative phosphotransferase, containing a nucleotide (possibly ATP) binding motif. There have been some evidences of phosphoenolpyruvate phosphotransferase (PTS) systems being involved in biofilm formation in Vibrio cholerae, E. coli and Streptococcus gordonii (Houot and Watnick, 2008;Lazazzera, 2010;Houot et al, 2010). YP_001085191, when searched against RefSeq (Pruitt et al, 2000) database, comes to be A. baumannii poly-beta-1, 6 N-acetyl-D-glucosamine export porin PgaA, which could be involved in the export of PGA to the cell exterior.…”

Expression, purification, crystallization and preliminary X-ray analysis of the EIICGlcdomain of theEscherichia coliglucose transporter

“…In addition to carbohydrate uptake, the PTS also controls carbon and nitrogen metabolism, chemotaxis, biofilm formation and other bacterial behaviours (Lux et al, 1995(Lux et al, , 1999Erni, 2006;Deutscher et al, 2006;Houot & Watnick, 2008;Poncet et al, 2009). The PTS is at the heart of a regulatory network known as carbon catabolite repression (Gorke & Stulke, 2008).…”