CRISPR-Cas protein Cas3 controls virulence in the oral pathogen Porphyromonas gingivalis.
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ABSTRACT:
ORGANISM(S): Galleria mellonella Porphyromonas gingivalis
PROVIDER: GSE154569 | GEO | 2020/09/16
REPOSITORIES: GEO
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Project description:CRISPR-Cas protein Cas3 controls virulence in the oral pathogen Porphyromonas gingivalis.
| PRJNA646710 | ENA
Project description:Porphyrmonas gingivalis is an oral pathogen associated with the inflammatory disease periodontitis. The colonization of oral epithelial surfaces by P. gingivalis may also lead to the autoimmune disease rheumatoid arthritis. One of the hallmarks of rheumatoid arthritis is the loss of tolerance against citrullinated proteins. Citrullination is a post-translational modification of arginine residues, leading to a change in structure and function of the respective protein. This modification is catalysed by peptidylarginine deiminases (PAD). Interestingly, P. gingivalis secretes a citrullinating enzyme, known as P. gingivalis PAD (PPAD), which targets bacterial and human proteins. Since the extent of P. gingivalis protein citrullination by PPAD was not yet known, the present study was aimed at identifying the extracellular proteome and citrullinome of P. gingivalis. To this end, extracellular proteins of two reference strains, two PPAD-deficient mutants and three clinical isolates of P. gingivalis were analysed by mass spectrometry. The results uncovered substantial heterogeneity in the extracellular proteome and citrullinome of P. gingivalis, especially in relation to the extracellular detection of typical cytoplasmic proteins. In contrast, major virulence factors were identified in all investigated isolates although their citrullination was shown to vary. This may be related to post-translational processing of the PPAD enzyme.
2018-02-28 | PXD003444 | Pride
Project description:Mechanisms by which Porphyromonas gingivalis (P. gingivalis) infection enhances oral tumor growth or resistance to cell death remain elusive. Here, we determined that P. gingivalis infection mediates therapeutic resistance via inhibiting lethal mitophagy in cancer cells and tumors. Mechanistically, P. gingivalis targets the LC3B-ceramide complex by associating with LC3B via bacterial major fimbriae (FimA) protein, preventing ceramide-dependent mitophagy in response to various therapeutic agents. Moreover, ceramide-mediated mitophagy is induced by Annexin A2 (ANXA2)-ceramide association, involving the E142 residue of ANXA2. Inhibition of ANXA2-ceramide-LC3B complex formation by wild-type P. gingivalis prevented ceramide-dependent mitophagy. Moreover, a FimA-deletion mutant P. gingivalis variant had no inhibitory effects on ceramide-dependent mitophagy. Further, 16S rRNA sequencing of oral tumors indicated that P. gingivalis infection altered the microbiome of the tumor macroenvironment in response to ceramide analog treatment in mice. Thus, these data provide a mechanism describing the pro-survival roles of P. gingivalis in oral tumors.
2024-06-23 | PXD051568 | Pride
Project description:The CRISPR (clustered regularly interspaced short palindromic repeat)-Cas system is a unique genomic entity that provides prokaryotic cells with adaptive and heritable immunity. Initial studies identified CRISPRs as central elements used by bacteria to protect against foreign nucleic acids; however, emerging evidence points to CRISPR involvement in bacterial virulence. The present study aimed to identify the participation of one CRISPR-Cas protein, Cas3, in the virulence of the oral pathogen Porphyromonas gingivalis, an organism highly associated with periodontitis. Our results show that compared to the wild type, a mutant with a deletion of the Cas3 gene, an essential nuclease part of the class 1 type I CRISPR-Cas system, increased the virulence of P. gingivalis In vitro infection modeling revealed only mildly enhanced production of proinflammatory cytokines by THP-1 cells when infected with the mutant strain. Dual transcriptome sequencing (RNA-seq) analysis of infected THP-1 cells showed an increase in expression of genes associated with pathogenesis in response to Δcas3 mutant infection, with the target of Cas3 activities in neutrophil chemotaxis and gene silencing. The importance of cas3 in controlling virulence was corroborated in a Galleria mellonella infection model, where the presence of the Δcas3 mutant resulted in a statistically significant increase in mortality of G. mellonella A time-series analysis of transcription patterning during infection showed that G. mellonella elicited very different immune responses to the wild-type and the Δcas3 mutant strains and revealed a rearrangement of association in coexpression networks. Together, these observations show for the first time that Cas3 plays a significant role in regulating the virulence of P. gingivalis IMPORTANCE Porphyromonas gingivalis is a key pathogen of periodontitis, a polymicrobial disease characterized by a chronic inflammation that destroys the tissues supporting the teeth. Thus, understanding the virulence potential of P. gingivalis is essential to maintaining a healthy oral microbiome. In nonoral organisms, CRISPR-Cas systems have been shown to modulate a variety of microbial processes, including protection from exogenous nucleic acids, and, more recently, have been implicated in bacterial virulence. Previously, our clinical findings identified activation of the CRISPR-Cas system in patient samples at the transition to disease; however, the mechanism of contribution to disease remained unknown. The importance of the present study resides in that it is becoming increasingly clear that CRISPR-associated proteins have broader functions than initially thought and that those functions now include their role in the virulence of periodontal pathogens. Studying a P. gingivalis cas3 mutant, we demonstrate that at least one of the CRISPR-Cas systems is involved in the regulation of virulence during infection.
| S-EPMC7527141 | biostudies-literature
Project description:Recent epidemiological studies revealed a significant association between oral squamous cell carcinoma (OSCC) and Porphyromonas gingivalis, a major pathogen of periodontal disease. As a keystone pathogen, P. gingivalis is known not only to damage local periodontal tissue, but also to evade from the host immune system and eventually affect systemic health. However, its role in OSCC has yet to be well studied. To explore the underlying effect of chronic P. gingivalis infection on OSCC and to identify relevant biomarkers as promising targets for therapy and prevention, we established a novel model by exposing human immortalized oral epithelial cells (HIOECs) to P. gingivalis at a low MOI for 5 to 23 weeks. The P. gingivalis-infected HIOECs were monitored in tumor biological alteration. Bioinformatics analyses were performed on HIOECs infected for 15 weeks, and some selected data were validated by q-PCR and (or) Western blot on cells infected for 15 and 23 weeks. Accordingly, we found that persistent exposure to P. gingivalis induced tumor biological properties on HIOECs. Tumor-related genes such as NNMT, FLI1, GAS6, lncRNA CCAT1, PDCD1LG2 and CD274 were aberrantly expressed in response to long-term exposure of P. gingivalis. In addition, some clinical biomarkers and novel proteins were presented. In conclusion, chronic P. gingivalis infection may be a potential risk factor of OSCC. The key regulators and biomarkers might be used in monitoring OSCC with chronic periodontal infection. We applied lncRNA Microarray (Affymetrix) which could simultaneously detect coding and non-coding genes to explore the differences of cell gene expression after long-term exposure to P. gingivalis.
2017-06-01 | GSE87539 | GEO
Project description:High-density tiling microarray and RNA sequencing technologies were used to analyze the transcriptome of Porphyromonas gingivalis. The compiled P. gingivalis transcriptome were based on total RNA samples isolated from three different laboratory culturing conditions and the strand-specific transcription profiles generated covered the entire genome including both protein coding and non-coding regions. The transcription profiles revealed various operon structures, 5' and 3' end untranslated regions (UTRs), differential expression patterns, and several not-yet annotated transcripts within intergenic and antisense regions. Further transcriptome analysis identified the majority of the genes to be expressed within operons and most 5' and 3' ends to be protruding UTRs of which several 3M-CM-"M-BM-^@M-BM-^Y UTRs were extended to overlap genes encoded on the opposite/antisense strand. Extensive antisense RNAs were identified opposite to most of insertion sequence (IS) elements. With the growing realization that non-coding RNAs play important biological functions, the comprehensive transcriptome profiles compiled in this study are of great value for the further understanding of gene regulation and virulence mechanism in this periodontal pathogen. The transcriptome profiles can be viewed at and downloaded from the 'Microbial Transcriptome Database' web site http://bioinformatics.forsyth.org/mtd. 3 total RNA samples from growing P. gingivalis strain W83 under different media conditions subjected to RNA-Seq.
2011-10-01 | E-GEOD-30452 | biostudies-arrayexpress
Project description:Genotyping studies suggest that there is genetic variability among P. gingivalis strains, however the extent of variability remains unclear, and the regions of variability have only partially been identified. We previously used heteroduplex analysis of the ribosomal operon intergenic spacer region (ISR) to type P. gingivalis strains in several diverse populations, identifying 6 predominant heteroduplex types and many minor ones. In addition we used ISR sequence analysis to determine the relatedness of P. gingivalis strains to one another, and demonstrated a link between ISR sequence phylogeny and the disease-associated phenotype of P. gingivalis strains. The availability of whole genome microarrays based on the genomic sequence of strain W83 has allowed a more comprehensive analysis of P. gingivalis strain variability, using the entire genome. The objectives of this study were to define the phylogeny of P. gingivalis strains using the entire genome, to compare the phylogeny based on genome content to the phylogeny based on a single locus (ISR), and to identify genes that are associated with the strongly disease-associated strain W83 that could be important for virulence. Keywords: Comparative genomic hybridization
2008-10-09 | GSE13128 | GEO
Project description:High-density tiling microarray and RNA sequencing technologies were used to analyze the transcriptome of Porphyromonas gingivalis. The compiled P. gingivalis transcriptome were based on total RNA samples isolated from three different laboratory culturing conditions and the strand-specific transcription profiles generated covered the entire genome including both protein coding and non-coding regions. The transcription profiles revealed various operon structures, 5' and 3' end untranslated regions (UTRs), differential expression patterns, and several not-yet annotated transcripts within intergenic and antisense regions. Further transcriptome analysis identified the majority of the genes to be expressed within operons and most 5' and 3' ends to be protruding UTRs of which several 3â UTRs were extended to overlap genes encoded on the opposite/antisense strand. Extensive antisense RNAs were identified opposite to most of insertion sequence (IS) elements. With the growing realization that non-coding RNAs play important biological functions, the comprehensive transcriptome profiles compiled in this study are of great value for the further understanding of gene regulation and virulence mechanism in this periodontal pathogen. The transcriptome profiles can be viewed at and downloaded from the 'Microbial Transcriptome Database' web site http://bioinformatics.forsyth.org/mtd.
2011-10-01 | GSE30452 | GEO
Project description:Genotyping studies suggest that there is genetic variability among P. gingivalis strains, however the extent of variability remains unclear, and the regions of variability have only partially been identified. We previously used heteroduplex analysis of the ribosomal operon intergenic spacer region (ISR) to type P. gingivalis strains in several diverse populations, identifying 6 predominant heteroduplex types and many minor ones. In addition we used ISR sequence analysis to determine the relatedness of P. gingivalis strains to one another, and demonstrated a link between ISR sequence phylogeny and the disease-associated phenotype of P. gingivalis strains. The availability of whole genome microarrays based on the genomic sequence of strain W83 has allowed a more comprehensive analysis of P. gingivalis strain variability, using the entire genome. The objectives of this study were to define the phylogeny of P. gingivalis strains using the entire genome, to compare the phylogeny based on genome content to the phylogeny based on a single locus (ISR), and to identify genes that are associated with the strongly disease-associated strain W83 that could be important for virulence. Keywords: Comparative genomic hybridization Comparative genomic analysis of 7 clinically prevalent P. gingivalis strains was performed, using whole genome microarrays based on the sequence of strain W83. Strain W83 was the reference strains and there were 6 test strains. Flip-dye replicates were performed.
2010-05-18 | E-GEOD-13128 | biostudies-arrayexpress
Project description:Wild type Porphyromonas gingivalis strain ATCC33277 (V3176) and PG1626 - deficient mutant (V3177) were grown in iron replete conditions was used to compare to Porphyromonas gingivalis strains grown in iron chelated conditions.
2023-06-08 | GSE174493 | GEO