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Gene regulation by the RcsCB two-component system and the auxiliary protein TviA in S. Typhi

ABSTRACT: In response to osmolarity, Salmonella enterica serotype Typhi (S. Typhi) regulates genes required for Vi capsular antigen expression oppositely to those required for motility and invasion. Previous studies suggest that osmoregulation of motility, invasion and capsule expression is mediated through the RcsC/RcsD/RcsB phosphorelay system. Here we performed gene expression profiling and functional studies to determine the role of TviA, an auxiliary protein of the RcsB response regulator, in controlling virulence gene expression in S. Typhi. TviA repressed expression of genes encoding flagella and the invasion associated type III secretion system (T3SS-1) through repression of the flagellar regulators flhDC and fliZ, resulting in reduced invasion and reduced expression of FliC. Both RcsB and TviA reduced expression of flhDC, but only TviA altered flhDC expression in response to osmolarity. These data suggest that the auxiliary TviA protein integrates a new regulatory input into the RcsB regulon of S. Typhi, thereby altering expression of genes encoding flagella, the Vi antigen and T3SS-1 in response to osmolarity.

ORGANISM(S): Salmonella enterica Salmonella enterica subsp. enterica serovar Typhi

PROVIDER: GSE15752 | GEO | 2009/12/11

SECONDARY ACCESSION(S): PRJNA116777

REPOSITORIES: GEO

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Gene regulation by the RcsCB two-component system and the auxiliary protein TviA in S. Typhi

Project description:In response to osmolarity, Salmonella enterica serotype Typhi (S. Typhi) regulates genes required for Vi capsular antigen expression oppositely to those required for motility and invasion. Previous studies suggest that osmoregulation of motility, invasion and capsule expression is mediated through the RcsC/RcsD/RcsB phosphorelay system. Here we performed gene expression profiling and functional studies to determine the role of TviA, an auxiliary protein of the RcsB response regulator, in controlling virulence gene expression in S. Typhi. TviA repressed expression of genes encoding flagella and the invasion associated type III secretion system (T3SS-1) through repression of the flagellar regulators flhDC and fliZ, resulting in reduced invasion and reduced expression of FliC. Both RcsB and TviA reduced expression of flhDC, but only TviA altered flhDC expression in response to osmolarity. These data suggest that the auxiliary TviA protein integrates a new regulatory input into the RcsB regulon of S. Typhi, thereby altering expression of genes encoding flagella, the Vi antigen and T3SS-1 in response to osmolarity. Gene expression profiles of the S. Typhi wild-type strain, a rcsB mutant, a (delta)tviB-vexE (tviBCDEvexABCDE) mutant, and a (delta)viaB (tviABCDEvexABCDE) mutant (lacking tviA expression) was compared. Strains were cultured in SOB broth (low salt), a condition known to induce expression of TviA and the RcsCB system. Total RNA was extracted from two independent cultures (biological replicates) of each strain. cDNA was differentially labeled with Alexa Fluor 555 and 648, respectively and competitively hybridized on S.Typhimurium arrays (PFGRC, version 4). The submitted samples are biological replicates of a competitive hybridization of cDNA from the wild-type strain (Ty2) and the rcsB mutant (SW237) (GSM395580, GSM395581) or the viaB mutant (STY2) and the tviB-vexE mutant (SW74) (GSM395582, GSM395583), respectively.

2010-05-18 | E-GEOD-15752 | biostudies-arrayexpress

Structure-based analysis of the gene network controlled by the Salmonella RcsB response regulator

Project description:RcsB is a transcriptional regulator that controls expression of numerous genes in Enterobacteriaceae. RcsB accomplishes this role alone or in combination with auxiliary transcriptional factors independently or dependently of phosphorylation. To understand the mechanisms by which RcsB regulates such a large number of genes, we performed structural and in vitro and in vivo functional studies with different RcsB variants. Our structural data reveal that RcsB binds the promoters of rprA and flhDC genes in a dimeric active conformation, docking the DNA-binding domains into the major groove of the DNA that preform a weak read-out of the target sequence. Interestingly, comparative structural analysis shows that DNA binding may trigger the active conformation in unphosphorylated RcsB. Furthermore, RNAseq analysis performed in strains expressing wild-type or several RcsB variants provided new insights into the contribution of phosphorylation to gene regulation and assign a potential role of RcsB in regulating iron and molybdenum metabolism. Finally, we delimited the RcsB box for homodimeric active RcsB binding with sequence TN(G/A)GAN4TC(T/C)NA that can be found in promoter and intragenic regions to regulate activation or inhibition of gene transcription.

2020-07-18 | GSE154668 | GEO

A comparison between two Escherichia coli K-12 MG1655 substrains possessing different swimming motility

Project description:Substrains in Escherichia coli K-12 MG1655 can possess various swimming motility, which is mostly resulted from different expression levels of flhDC. Here, we studied the swimming motility of two MG1655 substrains, CY562 and CY570. Our results showed that CY562 had no insertion at the promoter region of flhDC and possessed no swimming motility. In contrast, CY570 had an IS-element insertion at the promoter region of flhDC and showed a hyper-motile phenotype. Transcriptomic data suggest that expression of flhDC and the other known flagella genes was much lower in CY562 than that in CY570. Moreover, CY562 possessed higher expression levels for genes involved in stress response, especially acid-stress response, than CY570. Consistently, CY562 showed a higher survival rate under acid stress than CY570. Our data indicate that there are mechanisms conversely regulating motility and stress response in E. coli.

2021-11-15 | GSE165438 | GEO

Transcriptomic analysis reveals the gene expression profiles and cellular functions regulated by the RcsB in Yersinia enterocolitica

Project description:A total of 132 differentially expressed genes (DEGs) regulated by the RcsB have been identified, of which 114 were up-regulated and 18 were down-regulated. Analyses of the GO annotation and KEGG enrichment led us to focus on the role of RcsB in the inhibition of chemotactic behavior, flagella biosynthesis and bacterial infection in Y. enterocolitica. Overall, the information provided in this study complements our understanding of Rcs phosphorelay in the regulation of Y. enterocolitica pathogenicity, and, simultaneously, provides clues to additional roles of the Rcs system in other members of family Enterobacteriaceae.

2020-04-27 | GSE149365 | GEO

Salmonella typhimurium igaA(T191P) and related

Project description:We have exploited a spontaneously isolated mutant IgaA(T191P) that is near-maximally activated for the Rcs system, to identify a vast set of genes that respond, and report new regulatory properties of this signaling system in Salmonella enterica serovar Typhimurium. Microarray data show that the Rcs system normally functions as a positive regulator of SPI-2 and other genes important for growth of Salmonella in macrophages, although when highly activated, the system completely represses the SPI-1/SPI-2 virulence, flagellar, and fimbrial biogenesis pathways. The auxilliary protein RcsA, which works with RcsB to positively regulate colanic acid and other target genes, not only stimulates but also antagonizes the positive regulation of many genes in the igaA mutant. We show that RcsB represses motility through the 'RcsB box' in the promoter region of the master operon flhDC, and that RcsA is not required for this regulation. Curiously, RcsB selectively stimulates expression of the flagellar Type 3 secretion genes fliPQR; an RcsAB box located downstream of fliR influences this regulation. We show that excess colanic acid impairs swimming and inhibits swarming motility, consistent with the inverse regulation of the two pathways by the Rcs system. The work was published in Journal of Bacteriology: Wang Q, Zhao Y, McClelland M, Harshey RM. The RcsCDB signaling system and swarming motility in Salmonella enterica serovar Typhimurium: dual regulation of flagellar and SPI-2 virulence genes. J Bacteriol. 2007 Sep 28; [Epub ahead of print] PMID: 17905992 [PubMed - as supplied by publisher] Keywords: Comparative genomic hybridization Microarray was used to reveal the gene expression profiles in igaA(T191P) and related strains at two growth stages (log and stationary phases). Quantitative RT-PCR was used for detail studies.

2008-06-24 | E-GEOD-9584 | biostudies-arrayexpress

Salmonella typhimurium igaA(T191P) and related

2008-06-25 | GSE9584 | GEO

The response of Extra-intestinal Pathogenic Escherichia coli to human serum

Project description:In this study the transcriptional response of an ExPEC E. coli strain (CFT073) to human serum was investigated. In response, CFT073 up-regulated expression of iron and manganese acquisition systems and induced expression of iron regulated genes. High osmolarity of serum induced the osmotic shock response genes, promoting uptake of osmoprotectants by CFT073. Resistance of CFT073 to the bactericidal properties of serum involved increased expression of envelope stress regulators including CpxR, ?E and RcsB. Many of the up-regulated genes induced by active serum were regulated by the Rcs two component system. This system is triggered by envelope stress such as changes to cell wall integrity. RcsB-mediated serum resistance was conferred through induction of the exopolysaccharide colanic acid. Production of this exopolysaccharide may be protective while cell wall damage caused by serum components is repaired. Experimental Design: Two experiments are reported: 1) . The transcriptome of E. coli CFT073 exposed to LB supplemented with 50 % normal human serum was compared to that of bacteria grown in LB alone for 45 min, and 2) The transcriptome of CFT073 in response to normal healthy serum and heat inactivated serum (which has no bactericidal activity). Four biological replicates were performed per experiment with Dye swaps performed on sample replicates to eliminate any dye bias

2013-10-20 | E-MTAB-1985 | biostudies-arrayexpress

Identification of differentially expressed microRNAs between human HCC tumors with and without vascular invasion [batch 2]

Project description:Non-coding microRNAs (miRNAs) mainly regulate the expression of targeted genes by regulating mRNA degradation or repressing their protein translation. Recent research has shown miRNAs to have remarkable potential as biomarkers for the diagnosis and prognosis of disease. Since vascular invasion (VI) provides a direct path for solid tumor metastasis/recurrence, in this study we investigated if miRNAs can serve as a biomarker to differentiate between VI- and VI+ tumors. MiRNA microarray profiling was then performed on 172 human HCC tumors samples with or without VI from HCC patients.

2015-03-23 | GSE67139 | GEO

Identification of differentially expressed microRNAs between human HCC tumors with and without vascular invasion [batch 1]

2015-03-23 | GSE67138 | GEO

Proteus mirabilis mrpJ microarrays

Project description:This series of microarrays compares gene expression by the bacterial pathogen Proteus mirabilis when the transcriptional regulator mrpJ is deleted or induced to levels found during experimental urinary tract infection. The enteric bacterium Proteus mirabilis is associated with a significant number of catheter-associated urinary tract infections. Strict regulation of the antagonistic processes of adhesion and motility, mediated by fimbriae and flagella, respectively, is essential for successful disease progression. Previously, the transcriptional regulator MrpJ, which is encoded by the mrp fimbrial operon, has been shown to repress both swimming and swarming motility. Here we show that MrpJ affects a wide array of cellular processes beyond adherence and motility. Microarray analysis found that expression of mrpJ mimicking expression levels that occur during UTI leads to differential expression of 217 genes related to, among others, bacterial virulence, type VI secretion and metabolism. We probed the molecular mechanism of transcriptional regulation through MrpJ using reporter assays and chromatin immunoprecipitation (ChIP). Two virulence-associated target genes, the flagellar master regulator flhDC and mrp itself, appear to be regulated through a binding site proximal to the transcriptional start, complemented by a more distantly situated enhancer site. Furthermore, an mrpJ deletion mutant colonized the bladders of mice at significantly lower levels in a transurethral model of infection. Additionally, we observe that mrpJ is widely conserved in a collection of recent clinical isolates, leading us to conclude that our results elucidate an unanticipated role of MrpJ as a global regulator of P. mirabilis virulence. Four biological replicates were analyzed for each set of arrays (P. mirabilis HI4320 wild type vs. ΔmrpJ, and vector pLX3607 vs. mrpJ plasmid pLX3805).

2015-04-13 | E-GEOD-63321 | biostudies-arrayexpress

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