Chemotaxis Gene Expression Research Articles

N-acyl-homoserine lactones-mediated quorum sensing promotes intestinal colonization of Aeromonas veronii through facilitating cheA gene expression

Aeromonas veronii, which is a dominant bacterial species that causes economically adverse diseases in aquaculture, can colonize the gastrointestinal tracts of humans and animals, causing severe infections. Preceding studies have demonstrated that N-acyl-homoserine lactones (AHLs)-mediated quorum sensing (QS) stimulates the intestinal colonization of A. veronii, however, the mechanism involved remains unclear. Here, we performed transcriptomics analyses of the wild-type strain A. veronii Z12 and the AHLs synthase gene (ahyI) mutant strain Z12ΔahyI. The results suggested that the AHLs-mediated QS system regulated about 254 differentially expressed genes (DEGs), which contained 82 up-regulated and 172 down-regulated DEGs in strain Z12ΔahyI. Functional annotation and enrichment analyses of the DEGs revealed that the AHLs-mediated QS system primarily regulated chemotaxis gene expression. According to qRT-PCR validation and chemotaxis measurements, the deletion of the ahyI gene lowered the expression of chemotaxis genes, as well as the chemotaxis ability. Among the chemotaxis genes, the sensory histidine kinase gene cheA was determined as the critical gene to investigate as relates to its effects on intestinal colonization. The cheA gene mutant strain Z12ΔcheA was constructed, and it was determined that the expression levels of chemotaxis genes and chemotaxis ability of strain Z12ΔcheA were significantly lowered, as well as the intestinal ecological niche competition of Z12ΔcheA with intestinal bacteria. Furthermore, strain Z12ΔcheA exhibited lower dispersion and colonization levels in the intestine, in contrast to A. veronii Z12. Our findings confirmed that the regulation of the AHLs-mediated QS system on expression of the cheA gene that can contribute to colonizing the host with A. veronii Z12, which can further elucidate pathogenic colonization strategies.

Comparative genomics of Geobacter chemotaxis genes reveals diverse signaling function

BackgroundGeobacter species are δ-Proteobacteria and are often the predominant species in a variety of sedimentary environments where Fe(III) reduction is important. Their ability to remediate contaminated environments and produce electricity makes them attractive for further study. Cell motility, biofilm formation, and type IV pili all appear important for the growth of Geobacter in changing environments and for electricity production. Recent studies in other bacteria have demonstrated that signaling pathways homologous to the paradigm established for Escherichia coli chemotaxis can regulate type IV pili-dependent motility, the synthesis of flagella and type IV pili, the production of extracellular matrix material, and biofilm formation. The classification of these pathways by comparative genomics improves the ability to understand how Geobacter thrives in natural environments and better their use in microbial fuel cells.ResultsThe genomes of G. sulfurreducens, G. metallireducens, and G. uraniireducens contain multiple (~70) homologs of chemotaxis genes arranged in several major clusters (six, seven, and seven, respectively). Unlike the single gene cluster of E. coli, the Geobacter clusters are not all located near the flagellar genes. The probable functions of some Geobacter clusters are assignable by homology to known pathways; others appear to be unique to the Geobacter sp. and contain genes of unknown function. We identified large numbers of methyl-accepting chemotaxis protein (MCP) homologs that have diverse sensing domain architectures and generate a potential for sensing a great variety of environmental signals. We discuss mechanisms for class-specific segregation of the MCPs in the cell membrane, which serve to maintain pathway specificity and diminish crosstalk. Finally, the regulation of gene expression in Geobacter differs from E. coli. The sequences of predicted promoter elements suggest that the alternative sigma factors σ28 and σ54 play a role in regulating the Geobacter chemotaxis gene expression.ConclusionThe numerous chemoreceptors and chemotaxis-like gene clusters of Geobacter appear to be responsible for a diverse set of signaling functions in addition to chemotaxis, including gene regulation and biofilm formation, through functionally and spatially distinct signaling pathways.

Transcription Profiling of the Stringent Response in Escherichia coli

The bacterial stringent response serves as a paradigm for understanding global regulatory processes. It can be triggered by nutrient downshifts or starvation and is characterized by a rapid RelA-dependent increase in the alarmone (p)ppGpp. One hallmark of the response is the switch from maximum-growth-promoting to biosynthesis-related gene expression. However, the global transcription patterns accompanying the stringent response in Escherichia coli have not been analyzed comprehensively. Here, we present a time series of gene expression profiles for two serine hydroxymate-treated cultures: (i) MG1655, a wild-type E. coli K-12 strain, and (ii) an isogenic relADelta251 derivative defective in the stringent response. The stringent response in MG1655 develops in a hierarchical manner, ultimately involving almost 500 differentially expressed genes, while the relADelta251 mutant response is both delayed and limited in scope. We show that in addition to the down-regulation of stable RNA-encoding genes, flagellar and chemotaxis gene expression is also under stringent control. Reduced transcription of these systems, as well as metabolic and transporter-encoding genes, constitutes much of the down-regulated expression pattern. Conversely, a significantly larger number of genes are up-regulated. Under the conditions used, induction of amino acid biosynthetic genes is limited to the leader sequences of attenuator-regulated operons. Instead, up-regulated genes with known functions, including both regulators (e.g., rpoE, rpoH, and rpoS) and effectors, are largely involved in stress responses. However, one-half of the up-regulated genes have unknown functions. How these results are correlated with the various effects of (p)ppGpp (in particular, RNA polymerase redistribution) is discussed.

Cascade regulation of Caulobacter flagellar and chemotaxis genes

The assembly of a functional flagellum in the bacterium Caulobacter crescentus requires the protein products of approximately 30 genes expressed in a temporally discrete and spatially distinct manner. Our current understanding of this system has been limited by the fact that purified protein products are available for only about one-fifth of these genes. A genetically engineered transposon promoter probe, Tn5-VB32, containing a promoterless gene encoding neomycin phosphotransferase II (NPTase II) was used to generate a series of non-motile (fla −), kanamycin resistant strains of C. crescentus. These transcription-fusions allow the expression of NPTase II to be controlled by flagellar promoters, and thus questions of temporal regulation of flagellar genes can be addressed without the need to obtain purified protein products. The flagellar promoters accessed by Tn5-VB32 exhibited temporal regulation analogous to the known flagellar and chemotaxis gene products. The expression of NPTase II in these mutants is read from a chimeric mRNA that initiates in a chromosomal fla promoter and continues through the inserted NPTase II gene. Thus, temporal regulation is controlled by modulating either the initiation of transcription, or transcript turnover, at specific times in the cell cycle. Epistatic interactions between the genes accessed by the promoter probe and other flagellar loci were studied in double fla mutants generated by transducing the promoterprobe mutations into spontaneously derived second-site fla-mutant backgrounds. The synthesis of both natural fla gene products and the accessed NPTase II was assayed in these strains using antisera to purified components of the flagellum and to purified NPTase II. On the basis of these interactions, a trans-acting hierarchy of flagellar and chemotaxis gene expression is proposed.

Temporal and spatial control of flagellar and chemotaxis gene expression during Caulobacter cell differentiation.

Each Caulobacter cell division yields daughter cells that differ from one another both structurally and functionally. By focusing on the biogenesis of the polar flagellum and the proteins of the chemosensory system, several laboratories have now defined an extensive network of genes whose temporal expression is controlled in the predivisional cell. The differential turn-on of these genes contributes to the generation of asymmetry in the predivisional cell in that the products of these genes are targeted to specific cellular locations. To define the mechanisms that mediate this temporal and spatial control, fla genes whose products are not known were accessed by the insertion of transposon-carried drug resistance markers. The transposons were altered so that upon insertion into the chromosome, transcription fusions are formed in which the promoter regions of fla genes drive the expression of the downstream promoter-less drug resistance genes. Assays of the differential placement of the promoter-less drug resistance proteins (encoded within the interrupted fla genes) allow us to determine whether the positioning of the fla gene products is controlled by signal sequences in their proteins, by specific mRNA-targeting sequences in the 5'-regulatory regions of these genes, or by specific transcription from only one of the two newly replicated chromosomes in the predivisional cell.

Analysis of the pleiotropic regulation of flagellar and chemotaxis gene expression in Caulobacter crescentus by using plasmid complementation.

The biosynthesis of the single polar flagellum and the proteins that comprise the chemotaxis methylation machinery are both temporally and spacially regulated during the Caulobacter crescentus cell-division cycle. The genes involved in these processes are widely separated on the chromosome. The region of the chromosome defined by flaE mutations contains at least one flagellin structural gene and appears to regulate flagellin synthesis and flagellar assembly. The protein product of the adjacent flaY gene was found to be required to regulate the expression of several flagellin proteins and the assembly of a functional flagellum. We demonstrate here that each of these genes is also required for the expression of chemotaxis methylation genes known to map elsewhere on the chromosome. In order to study the regulation of these genes, plasmids were constructed that contain either an intact flaYE region or deletions in the region of flaY. These plasmids were mated into a wild-type strain and into strains containing various Tn5 insertion and deletion mutations and a temperature-sensitive mutation in the flaYE region. The presence of a plasmid containing the flaYE region allowed the mutant strains to swim and to exhibit chemotaxis, to synthesize increased amounts of the flagellins, to methylate their "methyl-accepting chemotaxis proteins" (MCPs), and to regain wild-type levels of methyltransferase activity. Chromosomal deletions that extend beyond the cloned region were not complemented by this plasmid. Plasmids containing small deletions in the flaY region failed to restore to any flaY or flaE mutants the ability to swim or to assemble a flagellar filament. When mated into a wild-type strain, plasmids bearing deletions in the flaY region were found to be recessive. The pleiotropic regulation of flagellin synthesis, assembly, and chemotaxis methylation functions exhibited by both the flaY and flaE genes suggest that their gene products function in a regulatory hierarchy that controls both flagellar and chemotaxis gene expression.