Gene expression of wild-type Porphyromnas gingivalis 33277 vs the transposon-mutant, J5-c5.
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PROVIDER: PRJNA504467 | ENA |
REPOSITORIES: ENA
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Project description:The P. gingivalis 33277 transposon mutant J5-c5, isolated in this study, was demonstrated to exhibit a lipid A deacylase phenotype. The mutant is unable to deacylate lipid A, which confers to it, in stark contrast to wild-type, an inability to evade the powerful TLR4 innate immune response. The mutant was shown to contain five transposons. In order to determine the affected gene responsible for this phenotype, we took the approach of comparing gene expression between wild-type and J5-c5 transposon mutant. RNA was extracted from three separate biological samples of each strain and RNAseq performed using Illumina’s HiSeq sequencing facility at the University of Washington Center for Precision Diagnostics. RNAseq data was analyzed closely following guidance outlined in Law et al (doi:10.12688/f1000research.9005.2).
2019-04-09 | GSE122289 | GEO
Project description:Porphyromonas gingivalis ATCC 33277
| PRJNA471021 | ENA
Project description:Porphyromonas gingivalis ATCC 33277 Transcriptome or Gene expression
| PRJNA270304 | ENA
Project description:Porphyromonas gingivalis is a major pathogen associated with the microbial biofilm-mediated disease chronic periodontitis. P. gingivalis has an obligate requirement for iron and protoporphyrin IX which it satisfies by transporting heme and iron liberated from the human host. The level of cellular iron in P. gingivalis affects the expression of a distinct iron-associated regulon of 64 genes and low iron invokes an iron sparing response. Iron homeostasis is usually mediated in Gram-negative bacteria at the transcriptional level by the Ferric Uptake Regulator (Fur). There is a single predicted P. gingivalis Fur superfamily orthologue named Har (heme associated regulator) that lacks the conserved metal binding residues found in other Fur orthologues. We show that Har binds both heme and ferrous iron resulting in a conformational change in the protein. Har was unable to complement the Escherichia coli H1780 fur mutant and there was no change in cellular metal content in a P. gingivalis Har mutant compared with the wild-type. The Har regulon of 44 genes is not predicted to play a role in iron homeostasis. Together these data indicated that Har does not regulate iron homeostasis in P. gingivalis. However, Har was required for heme-responsive biofilm development and its regulon overlapped P. gingivalis regulons previously identified after growth in heme limitation or as a homotypic biofilm. P. gingivalis is unique as an iron-dependent Gram-negative bacterium with a single heme-binding Fur superfamily orthologue, Har, that does not regulate iron homeostasis. Paired samples were compared on the same microarray using a two-colour system. A total of 6 paired microarray hybridizations were performed representing 6 biological replicates, where a balanced dye design was used, with the overall analysis including three microarrays where P. gingivalis 33277 samples were labeled with Cy3 and the paired ECR455 samples were labeled with Cy5 and three other microarrays where samples were labeled with the opposite combination of fluorophores.
2015-04-01 | E-GEOD-37099 | biostudies-arrayexpress
Project description:Porphyromonas gingivalis 33277 wild-type vs Fur orthologue Har isogenic mutant ECR455
| PRJNA158399 | ENA
Project description:The genomes of P. gingivalis strains 33277 and 381 are highly related phylogenetically. However, 33277 displays a reduced capacity to stimulate HEK cell TLR2-dependent signaling and THP-1 cell-dependent IL-1β production compared to 381, suggesting strain-specific differences in the expression of one or more bacterial immune-modulatory cell surface molecules. Genomic sequencing identified a single nucleotide polymorphism in the 33277 fimB allele (A>T), encoding a truncated FimB protein, relative to the 381 fimB allele. Gene exchange experiments indicated that the 33277 fimB allele contributes to the reduced immune-stimulatory capacity of this strain. Transcriptomic analyses determined that multiple genes related to carboxy-terminal domain (CTD) family proteins, including the gingipains, were upregulated in strain 33277 relative to strain 381. A gingipain substrate degradation assay confirmed that cell surface gingipain activity is higher in 33277; and an isogenic mutant strain deficient for the gingipains exhibited an increased capacity to activate TLR2 signaling and induce IL-1β production. Furthermore, 33277 and 381 isogenic mutant strains devoid of CTD cell surface proteins displayed increased immune-stimulatory capacities compared to the wild-type parental strains, confirming an immune-suppressive role for the gingipains. Collectively, our data indicate that the combination of an intact fimB allele and limited cell surface gingipain expression in strain 381 contributes to its relatively potent immune-stimulatory activity. Conversely, a defective fimB allele and high levels of cell surface gingipains reduce the capacity of strain 33277 to elicit host cell innate immune responses.
2019-09-30 | GSE128899 | GEO
Project description:This study investigated how P. gingivalis persisters interacted with human gingival epithelial cells (HGECs) and affected the gene expression of cytokines, and the related signaling pathways, via RNA sequencing. P. gingivalis (ATCC 33277) was treated with metronidazole (100 μg/ml) for 6 hours to generate P. gingivalis persisters (M-PgPs). HGECs with or without IL-1β pretreatment were stimulated by M-PgPs and P. gingivalis for 24 hours, and those without any treatments served as the blank control (BL).
2022-05-31 | GSE184777 | GEO
Project description:Investigation of whole genome gene expression level changes in Porphyromonas gingivalis ATCC 33277 treated with an anti-adhesive extract from Myrothamnus flabellifolia compared to the untreated strain. Aim: Identification of anti-adhesive plant extracts against cell surface binding of Porphyromonas gingivalis. Materials and Methods: Polyphenol-enriched extract from Myrothamnus flabellifolia (MF) traditionally used for periodontitis was tested for inhibition of P. gingivalis adhesion to KB cells by FACS, for influence on gingipain activity, hemagglutination and by microarray analysis for effects on the bacterial transcriptome. P. gingivalis-induced inflammation parameters were monitored by RT-PCR. Results: MF (100 µg/ml) reduced P. gingivalis adhesion/invasion about 50% by interacting with fimbriae and bacterial OMPs. Microarray analysis of MF-treated bacteria indicated up-regulation of genes involved in cell adhesion. As confirmed by RT-PCR, fimbrillin- and Arg-gingipain-encoding genes were upregulated by MF. On the protein level, inhibition (70%) of Arg-gingipain activity was observed, while the corresponding Lys-gingipain was hardly influenced. MF also inhibited hemagglutination. While exposure to P. gingivalis resulted in an increased expression of inflammation-related genes in KB cells, pretreatment of KB cells with MF evoked cytoprotective effects concerning IL-1β, IL-6, IL-8 and TNFα gene expression as well as IL-6 release rates. Conclusions: While being cytoprotective, MF exerts strong anti-adhesive effects against P. gingivalis. Thus, MF may be useful for the prevention of P. gingivalis-associated periodontal diseases. The chip study used total RNA recovered from two separate MF-treated and two separate untreated Porphyromonas gingivalis ATCC 33277 cultures. Each chip measured the expression level of 1,842 genes from P. gingivalis ATCC 33277 with thirteen 60-mer probes per gene, with three-fold technical redundancy.
2010-09-02 | E-GEOD-23939 | biostudies-arrayexpress
Project description:Porphyromonas gingivalis is a major pathogen associated with the microbial biofilm-mediated disease chronic periodontitis. P. gingivalis has an obligate requirement for iron and protoporphyrin IX which it satisfies by transporting heme and iron liberated from the human host. The level of cellular iron in P. gingivalis affects the expression of a distinct iron-associated regulon of 64 genes and low iron invokes an iron sparing response. Iron homeostasis is usually mediated in Gram-negative bacteria at the transcriptional level by the Ferric Uptake Regulator (Fur). There is a single predicted P. gingivalis Fur superfamily orthologue named Har (heme associated regulator) that lacks the conserved metal binding residues found in other Fur orthologues. We show that Har binds both heme and ferrous iron resulting in a conformational change in the protein. Har was unable to complement the Escherichia coli H1780 fur mutant and there was no change in cellular metal content in a P. gingivalis Har mutant compared with the wild-type. The Har regulon of 44 genes is not predicted to play a role in iron homeostasis. Together these data indicated that Har does not regulate iron homeostasis in P. gingivalis. However, Har was required for heme-responsive biofilm development and its regulon overlapped P. gingivalis regulons previously identified after growth in heme limitation or as a homotypic biofilm. P. gingivalis is unique as an iron-dependent Gram-negative bacterium with a single heme-binding Fur superfamily orthologue, Har, that does not regulate iron homeostasis.
2015-04-01 | GSE37099 | GEO
Project description:In this in vitro study, using DNA microarray we investigate the differential gene expression of Porphyromonas gingicalis ATCC 33277 when growing in the presence or in absence of its own monospecies biofilm.
2018-11-17 | GSE122623 | GEO