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Haemophilus ducreyi transcriptome during human infection

ABSTRACT: 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.

ORGANISM(S): [Haemophilus] ducreyi

SUBMITTER: Dharanesh Gangaiah

PROVIDER: E-GEOD-75236 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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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.

2015-11-21 | GSE75236 | GEO

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

H. ducreyi 35000HP grown in the presence or absence of FCS

Project description:Haemophilus ducreyi 35000HP was subjected to different in vitro growth conditions that resemble the in vivo host environment (presence of serum), in order to ascertain the overall changes in gene expression. This information might identify genes important for colonization and/or survival inside the host. H. ducreyi 35000HP was grown for 8 hours in Columbia broth in the presence or absence of FCS. After 8 hours total RNA was isolated and processed for DNA microarray analysis. This study includes four biological replicates (paired samples of bacteria grown in the presence/absence of FCS), one of which (Rep1) was selected for dye swap.

2010-05-18 | E-GEOD-13851 | biostudies-arrayexpress

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. 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.

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Global gene expression profile of Haemophilus ducreyi 35000HPcpxRpML125 relative to 35000HPcpxRpLS88

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Haemophilus ducreyi 35000HPfis relative to 35000HP

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Haemophilus ducreyi 35000HPhfq relative to 35000HP

Project description:The goal of this study was to compare the global trascription profile of a Haemophilus ducreyi hfq deletion mutant to that of the wild type parental strain. This study includes three biological replicates (paired samples of mutant and wild type strains), and all three were subjected to dye swap.

2014-03-07 | E-GEOD-44535 | biostudies-arrayexpress

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

Haemophilus ducreyi 35000HPcpxA gene expression relative to 35000HP

Project description:Comparative analysis of the global gene expression of a Haemophilus ducreyi 35000HP cpxA deletion mutant relative to the wild type strain After the cpxA mutant was generated, both strain were grown in Columbia Broth. After 8 hr of growth RNA wa isolated and processed for DNA microarray analysis. This study includes three biological replicates (paired samples), and all three were subjected to dye swap.

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