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RpoE-dependent transcriptomes of Haemophilus ducreyi

ABSTRACT: To potentially understand how H. ducreyi responds to membrane stress, here we defined RpoE-dependent genes using RNA-Seq. We identified 180 RpoE-dependent genes, of which 98% were upregulated; a major set of these genes encodes homologues of envelope maintenance and repair factors. We also identified and validated a putative RpoE promoter consensus sequence, which was enriched in the majority of RpoE-dependent targets. Comparison of RpoE-dependent genes to those controlled by CpxR showed that each transcription factor regulated a distinct set of genes. Given that RpoE activated a large number of genes encoding envelope maintenance and repair factors and CpxRA represses genes encoding envelope-localized proteins, these data suggest that RpoE and CpxRA appear to play distinct yet complementary roles in regulating envelope homeostasis in H. ducreyi. RNA of Haemophilus ducreyi 35000HP wild-type strain containing a rpoE inducible plasmid and wild-type strain containing a control plasmid were collected at 0 minutes, 5 minutes, and 10 minutes after induction in quadruplicate.

ORGANISM(S): [Haemophilus] ducreyi

SUBMITTER: Dharanesh Gangaiah

PROVIDER: E-GEOD-75238 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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RpoE-dependent transcriptomes of Haemophilus ducreyi

Project description:To potentially understand how H. ducreyi responds to membrane stress, here we defined RpoE-dependent genes using RNA-Seq. We identified 180 RpoE-dependent genes, of which 98% were upregulated; a major set of these genes encodes homologues of envelope maintenance and repair factors. We also identified and validated a putative RpoE promoter consensus sequence, which was enriched in the majority of RpoE-dependent targets. Comparison of RpoE-dependent genes to those controlled by CpxR showed that each transcription factor regulated a distinct set of genes. Given that RpoE activated a large number of genes encoding envelope maintenance and repair factors and CpxRA represses genes encoding envelope-localized proteins, these data suggest that RpoE and CpxRA appear to play distinct yet complementary roles in regulating envelope homeostasis in H. ducreyi.

2015-11-21 | GSE75238 | GEO

[Haemophilus] ducreyi

Project description:RpoE-dependent transcriptomes of Haemophilus ducreyi

| PRJNA302860 | ENA

Haemophilus ducreyi transcriptome during human infection

Project description:To better understand the molecular mechanisms underlying Haemophilus ducreyi infection in humans, here we determined the transcriptional profile of H. ducreyi in human lesions using RNA-Seq and compared it to that of in vitro growth. We were able to show that the in vivo transcriptome did not resemble that of in vitro growth. Compared to the inoculum, H. ducreyi harvested from pustules differentially expressed ~120 genes, of which 68 were upregulated. A large proportion of the upregulated genes encoded homologs of proteins involved in nutrient transport, alternative carbon pathways, growth arrest response, heat shock response, and DNA recombination. H. ducreyi upregulated few genes or operons (hgbA, flp-tad, and lspB-lspA2) required for human infection; expression of these genes is known to increase under nutrient stress. Homologs of several genes involved in anaerobic metabolism and ascorbate utilization were upregulated in vivo, suggesting that the organism is adjusting its metabolism to anaerobiosis in vivo. RNA from Haemophilus ducreyi infected pustules were collected from four volunteers and performed RNA-Seq.

2015-11-21 | E-GEOD-75236 | biostudies-arrayexpress

Transcriptomes of ΔrelAspoT and ΔdksA mutants of Haemophilus ducreyi in mid-log, transition, and stationary phase

Project description:To better understand the role of the (p)ppGpp-mediated stringent response in control of H. ducreyi virulence determinants, here we defined genes potentially regulated by either (p)ppGpp or DksA by using RNA-seq. We were able to show that loss of either (p)ppGpp or DksA resulted in dysregulation of multiple genes including several known virulence determinants. We also show that loss of (p)ppGpp or DksA resulted in differential expression of putative H. ducreyi small RNAs. RNA of Haemophilus ducreyi wildtype, relAspoT and dksA mutants were collected at mid-log, transition, and stationary phases of growth, in quadruplicate using stranded RNA -seq.

2015-06-04 | E-GEOD-67202 | biostudies-arrayexpress

Transcriptomes of 35000HP, ÎcpxA, ÎcpxR mutant strains of Haemophilus ducreyi in mid-log, transition, and stationary phase

Project description:To better understand the role of H. ducreyi CpxRA in controlling virulence determinants, here we defined genes potentially regulated by CpxRA by using RNA-Seq. Activation of CpxR by deletion of cpxA repressed nearly 70% of its targets, including seven established virulence determinants. Inactivation of CpxR by deletion of cpxR differentially regulated few genes and increased the expression of one virulence determinant. We identified a CpxR binding motif that was enriched in downregulated but not upregulated targets. These data reinforce the hypothesis that CpxA phosphatase activity plays a critical role in controlling H. ducreyi virulence in vivo. Characterization of the downregulated genes may offer new insights into pathogenesis. RNA of Haemophilus ducreyi wildtype, cpxA and cpxR mutants were collected at mid-log, transition, and stationary phases of growth, in quadruplicate.

2015-11-21 | E-GEOD-75237 | biostudies-arrayexpress

RpoE regulon in Rhodobacter sphaeroides

Project description:In Rhodobacter sphaeroides a transcriptional response to the reactive oxygen species singlet oxygen is controlled by the group IV sigma factor RpoE and the anti-sigma factor ChrR. In this study, we integrated various large datasets to identify genes within the singlet oxygen stress response that contain RpoE-dependent promoters within R. sphaeroides. Transcript pattern clustering and a RpoE-binding sequence model were used to predict candidate promoters that respond to singlet oxygen stress in R. sphaeroides. These candidate promoters were experimentally validated to nine R. sphaeroides RpoE-dependent promoters that control the transcription of 15 genes activated by singlet oxygen.

2010-07-01 | GSE22576 | GEO

RpoE regulon in Rhodobacter sphaeroides

2010-06-30 | E-GEOD-22576 | biostudies-arrayexpress

The single ECF sigma factor of Xylella fastidiosa is involved in the heat shock response

Project description:Genome sequence analysis of the bacterium Xylella fastidiosa revealed the presence of two genes, named rpoE and rseA, predicted to encode an ECF sigma factor and an anti-sigma factor, respectively. In this work, an rpoE null mutant was constructed in the citrus strain J1a12 and shown to be sensitive to exposure to heat shock and ethanol. To identify the X. fastidiosa σE regulon, global gene expression profiles were obtained by DNA microarray analysis of bacterial cells under heat shock identifying 23 sigmaE-dependent genes. Keywords: stress response, heat shock, rpoE mutant strain

2006-11-17 | GSE4960 | GEO

Determination of an Interaction Network between an Extracellular Bacterial Pathogen and the Human Host

Project description:A major gap in understanding infectious diseases is the lack of information about molecular interaction networks between pathogens and the human host. Haemophilus ducreyi causes the genital ulcer disease chancroid in adults and is a leading cause of cutaneous ulcers in children in the tropics. We developed a model in which human volunteers are infected on the upper arm with H. ducreyi until they develop pustules. To define the H. ducreyi and human interactome, we determined bacterial and host transcriptomic and host metabolomic changes in pustules. We found in vivo H. ducreyi transcripts were distinct from those in the inocula as were host transcripts in pustule and wounded control sites. Many of the upregulated H. ducreyi genes were involved in ascorbic acid and anaerobic metabolism and inorganic ion/nutrient transport. The top 20 significantly expressed human pathways showed that all were involved in immune responses. We generated a bipartite network for interactions between host and bacterial gene transcription; multiple positively correlated networks contained H. ducreyi genes involved in anaerobic metabolism and host genes involved with the immune response. Metabolomic studies showed that pustule and wounded samples had different metabolite compositions; the top ion pathway involved ascorbate and aldarate metabolism, which correlated with the H. ducreyi transcriptional response and upregulation of host genes involved in ascorbic acid recycling. These data show that an interactome exists between H. ducreyi and the human host and suggests that H. ducreyi exploits the metabolic niche created by the host immune response.

2019-05-30 | GSE130901 | GEO

The delta subunit of RNA polymerase, RpoE, is a global modulator of Streptococcus mutans environmental adaptation

Project description:Transcriptional profiling of S. mutans rpoE knockout mutant comparing to the wild type. The mutation of rpoE cause dramatic changes in phenotypes. The goal is to determine the genes affected by rpoE. The delta subunit of RNA polymerase, RpoE, is wide spread in low G+C Gram-positive bacteria and is thought to play a role in enhancing transcriptional specificity by blocking RNA polymerase binding at weak-promoter sites and stimulating RNA synthesis by accelerating core enzyme recycling. Despite the well-studied biochemical properties of RpoE, a role for this protein in vivo has not been defined in depth. In this study, we show that inactivation of rpoE in the human dental caries pathogen Streptococcus mutans causes impaired growth, and loss of important virulence traits, including biofilm formation, resistance to antibiotics, and tolerance to environmental stresses. Complementation of the mutant with rpoE expressed in trans restored its phenotype to wild type. The luciferase fusion reporter showed that rpoE was highly transcribed throughout growth, and that acid and hydrogen peroxide stresses repressed rpoE expression. Transcriptome profiling of wild type and ΔrpoE cells in the exponential and early stationary phase of growth, under acid and hydrogen peroxide stress and under both stresses combined revealed that genes involved in histidine synthesis, malolactic fermentation, biofilm formation, and antibiotic resistance were down-regulated in the ΔrpoE mutant in all conditions. Moreover, the loss of RpoE resulted in dramatic changes in transport and metabolism of carbohydrates and amino acids. Interestingly, differential expression, mostly up-regulation, of 330 non-coding regions was found. In conclusion, this study demonstrates that RpoE is an important global modulator of gene expression in S. mutans, which is required for optimal growth and environmental adaptation.

2010-09-21 | GSE22333 | GEO

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