Enterococcus faecalis EfaR mutant transcriptome profile
Ontology highlight
ABSTRACT:
ORGANISM(S): Enterococcus faecalis Enterococcus faecalis V583
PROVIDER: GSE33698 | GEO | 2012/06/19
SECONDARY ACCESSION(S): PRJNA148281
REPOSITORIES: GEO
Similar Datasets
Project description:Metal ions are important cofactors of inumerous enzymes and can interfere in many biological processes. Manganese is a particularly relevant metal as it is also involved in protection from oxidative stress and several Mn-dependent regulators have been shown to have a role in bacterial pathogenicity. A putative DNA-binding motif, here called efa motif was discovered in the promoter regions of genes previously shown to be differentially expressed in the presence of excess zinc, manganese and copper ions. This motif led to the finding and study of the genes putatively involved in the maintenance of manganese homeostasis in E. faecalis. In these genes is included the efaCBA operon, regulated by the Mn-dependent regulator EfaR. Other genes with this efa motif are shown to be regulated by EfaR. Regulation of these efa motif genes by EfaR is discussed as well as some clues on metal interaction with EfaR. Experiments mimicking conditions occurring on biological processes in the host showed that an EfaR plays an important role in biological processes crucial for E. faecalis colonization and virulence. Comparison of the transcriptome profiles of Enterococcus faecalis strains VE14089 and VE14089 efaR::tet 2 condition experiment: E. faecalis VE14089 cells vs. E. faecalis VE14089 efaR::tet cells
2012-06-18 | E-GEOD-33698 | biostudies-arrayexpress
Project description:We assessed changes in gene expression in response to manganese availability for the human pathogen Corynebacterium diphtheriae. Total RNA was harvested from wild-type C. diphtheriae strain 1737 and an isogenic ΔmntR (dip0619) grown in semi-defined metal-limited media (mPGT) without or with 5 µM manganese chloride supplementation. Three biological replicates were prepared and five genes were assessed by real-time PCR to validate the array results: dip0124, dip0169, dip0615, dip1923, and dip2261. (Abstract) Corynebacterium diphtheriae is the causative agent of a severe respiratory disease in humans. The bacterial systems required for infection are poorly understood, but the acquisition of metals such as manganese (Mn) are likely critical for host colonization. MntR is a Mn-dependent transcriptional regulator in C. diphtheriae and was previously shown to repress the expression of the mntABCD genes, which encode an ABC metal transporter. However, other targets of Mn and MntR regulation in C. diphtheriae have not been identified. In this study, we use comparisons between the gene expression profiles of wild-type C. diphtheriae strain 1737 grown without or with Mn supplementation and comparisons of gene expression between wild-type and an mntR mutant to characterize the C. diphtheriae Mn and MntR regulon. MntR was observed to both repress and induce the expression of various target genes in a Mn-dependent manner. Genes induced by MntR include the Mn-superoxide dismutase, sodA, and the putative ABC transporter locus, iutABCD. DNA binding studies showed that MntR interacts at the promoter regions for several genes identified in the expression study, and a 17-bp consensus MntR DNA binding site was identified. We found that an mntR mutant displayed increased sensitivity to Mn and Cd that could be alleviated by the additional deletion of the mntA-D transport locus, providing evidence that the MntABCD transporter functions as a Mn uptake system in C. diphtheriae. The findings in this study further our understanding of metal uptake systems and global metal regulatory networks in this important human pathogen.
2021-05-06 | GSE173960 | GEO
Project description:ICP-MS analysis of Streptococcus pneumoniae D39 reveals a high cell-associated manganese (Mn(II)) concentration that is comparable to that of zinc (Zn(II)). Stressing these cells with 100-200 µM Zn(II) leads to a slow-growth phenotype and a reduction in total Mn(II) concentration, with no decrease in the concentrations of other metal ions. Supplementation of the growth media with as little as 10 µM Mn(II) fully restores wildtype growth patterns and normal levels of cell-associated Mn(II). DNA microarray analysis reveals that zinc stress induces the expected upregulation of czcD (encoding a zinc effluxer), but also a pleiotropic transcriptional response suggestive of mild cell wall stress. Genes encoding a nitric oxide detoxification system (nmlR) and the Mn(II) uptake system (psaABC) are also induced. We conclude that Zn(II) toxicity results in a cytoplasmic Mn(II) deficiency, possibly caused by competition at the Mn(II) uptake transporter protein PsaA.
2011-02-02 | GSE23504 | GEO
Project description:Iron (Fe) and Manganese (Mn) are essential for bacterial survival and virulence as they are involved in catalysis, infection and biofilm formation. The basal metabolism of pathogenic bacteria would be disturbed by the deficiency of metal ions in the host environment. In this study, we found that Streptococcus pneumoniae (S. pneumoniae) can still grow slowly in the medium without manganese ions. Further ICP-MS determination showed that the iron concentration in the bacterium was increased when the manganese content was decreased. The supplement of iron in the manganese deficient culture medium (MDCM) can recovery the bacterial growth. The quantitative proteome using stable-isotope dimethyl labeling was performed to investigate the adaptive growth mechanism of S. pneumoniae in Mn-deficiency condition. Compared with the bacteria cultured in the normal medium, 25 proteins were down-regulated and 54 proteins were up-expressed with more than 1.2-fold alteration (P < 0.05) in S. pneumoniae cultured with MDCM. A large number of depressed proteins are participated in cell energy metabolism, amino acid synthesis process and oxidation products reduction process. The subsequent detection indicated that both intracellar and extracellular hydrogen peroxide levels of in cells were significantly elevated, and the intracellular ATP levels of were evidently decreased in cells cultured in the absence of manganese, meaning that Mn-deficiency can cause multiple metabolic disorders. More importantly, several iron ABC transporters, such as PiuA/PiaA/PitA/SPD_1609, etc, were over-expressed to uptake more iron from the medium. These results suggest that lacking of manganese only disturbed multiple metabolic processes of S. pneumoniae, but also caused the compensatory effect of iron for manganese Mn which is beneficial to the survival of bacteria in extreme environments.
2023-09-30 | PXD008652 | Pride
Project description:As transition metal availability is very limited inside the human host, fungal pathogens have evolved sophisticated mechanisms to uptake and utilize these micronutrients at the infection interface. While considerable attention was turned into iron, copper and zinc acquisition mechanisms and their importance in fungal fitness, less was done regarding either the role of Mn in infectious processes or the cellular mechanism by which fungal cells achieve their Mn-homeostasis. Here, we undertook a transcriptional profiling of the pathogenic fungus Candida albicans experiencing both Mn starvation and excess to comprehensibly capture biological processes that are modulated by Mn. We uncovered that Mn scarcity influence diverse biological processes associated with fungal fitness including morphogenetic switch, invasion of host cells and antifungal sensitivity. We also uncovered that Mn levels influence the abundance of iron and zinc emphasizing the complex crosstalk between ions metals. Deletion of SMF12, a member of Mn Nramp transporters confirmed its contribution to Mn uptake. In accordance with the RNA-seq data, smf12 was unable to form hyphae and damage host cells, and exhibited sensitivity to azole antifungals. We also found that unfolded protein response (UPR), likely activated by decreased glycosylation under Mn limitation, was essential to promote C. albicans growth. RNA-seq profiling of cells exposed to Mn excess uncovered that the UPR signaling was also essential to bypass Mn toxicity. Collectively, this study underscores the importance of Mn homeostasis in fungal virulence, and comprehensively provides a transcriptional portrait of biological functions that are modulated by Mn in a fungal pathogen.
2024-02-13 | GSE245114 | GEO
Project description:The human ZRT/IRT-like protein 8 (ZIP8), encoded by the solute carrier family 39 member 8 gene SLC39A8, is a metal cation symporter located mainly on the cell membrane. ZIP8 is well-known for its role in transporting divalent metal ions into the cells – these ions include several essential micronutrients (e.g., iron [Fe], manganese [Mn], and zinc [Zn]) and non-essential toxic heavy metal cadmium (Cd). In the current project, ZIP8 gene in the human cervical cancer HeLa cell line was knockout (KO) using the CRISPR/Cas9 genome editing technology. Then, the proteomes of ZIP8-KO HeLa cell line and HeLa parental cell line (with wildtype ZIP8 gene sequence) were quantified using iTRAQ and LC-MS/MS.
2022-10-20 | PXD036680 | Pride
Project description:Microarray analysis of Streptococcus pneumoniae TIGR4 transcriptome in response to manganese as the transcriptome changes in response to intracellular manganese accumulation via a mutation in sp1552/mntE a manganese efflux protein. Investigating role of manganese efflux and accumulation in S. pneumoniae: 3 TIGR4 in ThyB vs. TIGR4 in Mn and 3 TIGR4 in Mn vs mntE1 in Mn replicate 3
2010-05-22 | E-GEOD-14324 | biostudies-arrayexpress
Project description:The essential metal manganese (Mn) induces neuromotor disease at elevated levels. The manganese efflux transporter SLC30A10 regulates brain Mn levels. Homozygous loss-of-function mutations in SLC30A10 induce hereditary Mn neurotoxicity in humans. Our prior characterization of Slc30a10 knockout mice recapitulated the high brain Mn levels and neuromotor deficits reported in humans. But, mechanisms of Mn-induced motor deficits due to SLC30A10 mutations or elevated Mn exposure are unclear. To gain insights into this issue, we characterized changes in gene expression in the basal ganglia, the main brain region targeted by Mn, of Slc30a10 knockout mice using unbiased transcriptomics. Compared with littermates, >1,000 genes were upregulated or downregulated in the basal ganglia sub-regions (i.e., caudate putamen, globus pallidus, and substantia nigra) of the knockouts. Pathway analyses revealed notable changes in genes regulating synaptic transmission and neurotransmitter function in the knockouts that may contribute to the motor phenotype. Expression changes in the knockouts were essentially normalized by a reduced Mn chow, establishing that changes were Mn-dependent. Upstream regulator analyses identified hypoxia-inducible factor (HIF) signaling, which we recently characterized to be a primary cellular response to elevated Mn, as a critical mediator of the transcriptomic changes in the basal ganglia of the knockout mice. HIF activation was also evident in the liver of the knockout mice. These results: (1) enhance understanding of the pathobiology of Mn-induced motor disease; (2) identify specific target genes/pathways for future mechanistic analyses; and (3) independently corroborate the importance of the HIF pathway in Mn homeostasis and toxicity.
2024-02-09 | GSE247882 | GEO
Project description:Emerging antibiotic resistance among clinically relevant bacteria, paired with their ability to form biofilms on medical and technical devices, represents a serious problem in terms of effective and long-term decontamination in health care environments and gives rise to an urgent need for new antimicrobial materials. Here we present the first study of the impact of AGXX®, a novel broad-spectrum antimicrobial surface coating consisting of micro galvanic elements formed by silver and ruthenium, on the transcriptome of the nosocomial pathogen Enterococcus faecalis. E. faecalis was subjected to metal stress by growing it for different periods of time in the presence of AGXX® or silver-coated steel meshes. Subsequently, total RNA was isolated and next-generation RNA sequencing was performed to analyze variations in gene expression levels in the presence of the antimicrobial materials with focus on known stress genes. Exposure to AGXX® had a large impact on the transcriptome of E. faecalis. After 24 minutes almost 1/5 of the E. faecalis genome displayed differential expression. At each time-point the cop operon was strongly up-regulated, providing indirect evidence for the presence of free Ag+-ions. Moreover, exposure to AGXX® induced a broad general stress response in E. faecalis. Genes coding for the chaperones GroEL and GroES as well as the Clp proteases ClpE and ClpB were among the top up-regulated heat shock genes. Furthermore, differential expression of genes coding for thioredoxin, superoxide dismutase and glutathione synthetase indicates a high level of oxidative stress. We postulate a mechanism of action where the combination of Ag+-ions and reactive oxygen species generated by AGXX® results in a synergistic antimicrobial effect, which is superior to that of conventional silver coatings. Gene expression analysis of Enterococcus faecalis 12030 either subjected to metal stress by exposure to an antimicrobial AGXX®- or Ag-coated V2A steel mesh or exposed to an uncoated V2A steel mesh or left untreated performing RNA Sequencing with an Ion ProtonTM Sequencer and subsequent data analysis with a T-REx RNA-Sequencing expression analysis pipeline.
2016-03-16 | E-GEOD-79250 | biostudies-arrayexpress
Project description:Members of Acidobacteria phylum are difficult to isolate and grow. Nevertheless, for some isolated strains, trace elements solution is needed in culture media for Acidobacteria growth but the impact of these metals on their growth and metabolism is not known. Here we evaluated the effect of trace element solution SL-10 in on the growth of two strains (5B5 and WH15) of Acidobacteria belonging to Granulicella sp. and studied the proteomic responses to the metal with the best growth results, complementing with genomic characterization. Granulicella species had highest growth with the addition of Mn, as well as higher tolerance to this metal compared to seven other metal ions. Variations in tolerance to metal ion concentrations suggests that Granulicella sp. strains possess different mechanisms to deal with metal ion homeostasis and stress. The proteomic profiles of both strains indicated that Mn was more important in enhance enzymatic activity than to protein expression regulation. We did not find the most common transcriptional regulation of Mn homeostasis and we found candidate transporters that could be potentially involved in Mn homeostasis for Granulicella species. The presence of such transporters might be involved in tolerance to higher Mn concentrations, improving the adaptability of bacteria to metal enriched environments, such as the decaying wood-rich Mn environment where these two strains were isolated. Furthermore, strain 5B5 is likely more adapted to survive in an environment with higher concentration of several metal ions when compared to strain WH15.
2021-02-15 | PXD016551 | Pride