The phage-encoded PIT4 protein impacts the Pseudomonas aeruginosa type IV pili by affecting different two-component systems
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ABSTRACT:
PROVIDER: PRJNA962838 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR24333000_1.fastq.gz | Fastqsanger.gz | |||
| SRR24333000_2.fastq.gz | Fastqsanger.gz | |||
| SRR24333001_1.fastq.gz | Fastqsanger.gz | |||
| SRR24333001_2.fastq.gz | Fastqsanger.gz | |||
| SRR24333002_1.fastq.gz | Fastqsanger.gz |
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Project description:Two component systems (TCSs) control a large proportion of virulence factors in Pseudomonas aeruginosa. Yet, investigations on inhibitors of regulatory pathways of TCS remain scare, despite their potential in anti-virulence strategies. This work encompasses the working mechanism of PIT4, a protein derived from the lytic P. aeruginosa phage LSL4. This viral protein inhibits bacterial motility and in particular twitching motility, while reducing the virulence of P. aeruginosa towards HeLa cells. Via differential gene expression and a yeast two-hybrid screen, PIT4 was shown to interact with components of different two component systems. In one-on-one interaction assays, it was confirmed that PIT4 interacts with the histidine kinases FleS, PilS and PA2882, through interaction with the histidine kinase domain. As such, this work highlights the potential of previously unknown phage proteins in virulence regulation of multidrug resistant pathogens that might be exploited for anti-virulence strategies and biotechnological applications.
2023-09-25 | GSE230851 | GEO
Project description:Type IV pili (T4P) are prevalent, polymeric surface structures in pathogenic bacteria, making them ideal targets for effective vaccines. However, bacteria have evolved efficient strategies to evade type IV pili-directed antibody responses. Neisseria meningitidis are prototypical type IV pili-expressing Gram-negative bacteria responsible for life threatening sepsis and meningitis. This species has evolved several genetic strategies to modify the surface of its type IV pili based on recombination, phase variation and the presence of different alleles of genes involved in posttranslational modifications. This results in changes in pilin subunit amino acid sequence, nature of glycosylation and phosphoform modification, but how these different processes affect antibody binding at the structural level is still unknown. To explore this question, we provide cryo-electron microscopy structures of pili of different sequence types with sufficiently high resolution to visualize posttranslational modifications. We then generated nanobodies directed against type IV pili which alter pilus function in vitro and vivo; and determined their structures complexed with their pilus target. We also determined how the different types of pili surface modifications alter nanobody binding. These different structures shed light on the impressive complementarity between the different strategies used by bacteria to avoid antibody binding. Importantly, we also show that structural information can be used to make informed modifications in nanobodies as countermeasures to these immune evasion mechanisms.
2024-02-29 | PXD046119 | Pride
Project description:The phytopathogen Xylella fastidiosa produces two classes of pili, long type IV pili and short type I pili, which are involved in motility and adhesion. In this work, we have investigated the role of σ54 factor and its involvement in the regulation of fimbrial biogenesis in X. fastidiosa. An rpoN null mutant was constructed from citrus strain J1a12, and analyses of global gene expression profile by microarrays comparing the wild type and rpoN mutant strains showed that few genes exhibited differential expression. At least one gene, pilA1 (XF2542), which encodes the structural pilin protein of type IV fimbriae, showed decreased expression in the rpoN mutant whereas an operon encoding proteins of type I fimbriae were twofold more expressed in the mutant. Quantitative real time RT-PCR (qRT-PCR) analysis confirmed that pilA1 transcript was significantly reduced in the rpoN mutant. The transcriptional start site of pilA1 was determined by primer extension, and a canonical σ54-dependent promoter was found upstream of the start site. Genome sequence analysis of X. fastidiosa revealed five paralogues of pilA, but microarray and qRT-PCR data demonstrated that only the pilA1 transcript was significantly affected in the rpoN mutant. The rpoN mutant made more biofilm than the wild type strain and presented a cell-cell aggregative phenotype. These results indicate that σ54 regulates biofilm formation, probably via differential regulation of genes involved in type IV and type I fimbrial biogenesis. Keywords: rpoN mutant strain, pili
2007-10-22 | GSE8097 | GEO
Project description:Many bacteria express filaments called type IV pili on their surface, which are involved in motility (twitching), surface adhesion, biofilm formation and DNA uptake (natural transformation). Type IV pili are comprised of a helical assembly of repeating pilin subunits that allow both flexibility and strength. They are therefore powerful structures that enable bacterial proliferation and genetic adaptation, potentially leading to the development of pathogenicity and antibiotic resistance. They are also targets for drug development. By electron cryo-microscopy and mass spectrometry, we show that the bacterium Thermus thermophilus produces two forms of type IV pilus, differing in structure and protein composition. We have determined the structures of both and built atomic models, which reveal a new pilin, how the subunits assemble and their glycosylation patterns. We also delineate the roles of the two filaments in promoting twitching and natural transformation.
2020-03-09 | PXD017353 | Pride
Project description:The phytopathogen Xylella fastidiosa produces two classes of pili, long type IV pili and short type I pili, which are involved in motility and adhesion. In this work, we have investigated the role of Ï54 factor and its involvement in the regulation of fimbrial biogenesis in X. fastidiosa. An rpoN null mutant was constructed from citrus strain J1a12, and analyses of global gene expression profile by microarrays comparing the wild type and rpoN mutant strains showed that few genes exhibited differential expression. At least one gene, pilA1 (XF2542), which encodes the structural pilin protein of type IV fimbriae, showed decreased expression in the rpoN mutant whereas an operon encoding proteins of type I fimbriae were twofold more expressed in the mutant. Quantitative real time RT-PCR (qRT-PCR) analysis confirmed that pilA1 transcript was significantly reduced in the rpoN mutant. The transcriptional start site of pilA1 was determined by primer extension, and a canonical Ï54-dependent promoter was found upstream of the start site. Genome sequence analysis of X. fastidiosa revealed five paralogues of pilA, but microarray and qRT-PCR data demonstrated that only the pilA1 transcript was significantly affected in the rpoN mutant. The rpoN mutant made more biofilm than the wild type strain and presented a cell-cell aggregative phenotype. These results indicate that Ï54 regulates biofilm formation, probably via differential regulation of genes involved in type IV and type I fimbrial biogenesis. Direct comparison between wild type strain J1a12 and mutant strain rpoN-.
2010-06-25 | E-GEOD-8097 | biostudies-arrayexpress
Project description:DNA methylation is an important epigenetic mark in many eukaryotic organisms. De novo DNA methylation in plants can be achieved by the RNA-directed DNA methylation (RdDM) pathway, where the plant-specific DNA-dependent RNA polymerase Pol IV transcribes target sequences to initiate 24-nt siRNA production and action. The Arabidopsis DTF1/SHH1 has been shown to associate with Pol IV and is required for 24-nt siRNA accumulation and transcriptional silencing at several RdDM target loci. However, the extent and mechanism of DTF1 function in RdDM is unclear. We show here that DTF1 is necessary for the accumulation of the majority of Pol IV-dependent 24-nt siRNAs. It is also required for a large proportion of Pol IV-dependent de novo DNA methylation. Interestingly, there is a group of RdDM target loci where 24-nt siRNA accumulation but not DNA methylation is dependent on DTF1. Taken together, our results show that DTF1 is a core component of the RdDM pathway. Our results also suggest the involvement of DTF1 in an important negative feedback mechanism for DNA methylation at some RdDM target loci.
2013-06-03 | GSE44209 | GEO
Project description:DNA methylation is an important epigenetic mark in many eukaryotic organisms. De novo DNA methylation in plants can be achieved by the RNA-directed DNA methylation (RdDM) pathway, where the plant-specific DNA-dependent RNA polymerase Pol IV transcribes target sequences to initiate 24-nt siRNA production and action. The Arabidopsis DTF1/SHH1 has been shown to associate with Pol IV and is required for 24-nt siRNA accumulation and transcriptional silencing at several RdDM target loci. However, the extent and mechanism of DTF1 function in RdDM is unclear. We show here that DTF1 is necessary for the accumulation of the majority of Pol IV-dependent 24-nt siRNAs. It is also required for a large proportion of Pol IV-dependent de novo DNA methylation. Interestingly, there is a group of RdDM target loci where 24-nt siRNA accumulation but not DNA methylation is dependent on DTF1. Taken together, our results show that DTF1 is a core component of the RdDM pathway. Our results also suggest the involvement of DTF1 in an important negative feedback mechanism for DNA methylation at some RdDM target loci. Examination of whole-genome DNA methylation and small RNA in 12-day-old Col-0, dtf1-2, nrpd1-3 and nrpe1-11 seedlings
2013-06-03 | E-GEOD-44209 | biostudies-arrayexpress
Project description:In Myxococcus xanthus, two component systems have crucial roles in regulating motility behavior and development. Here, we describe an orphan response regulator consisting of an N-terminal receiver domain and a C-terminal DNA binding domain, which is required for A- and type IV pili dependent gliding motility. Genetic evidence suggests that phosphorylation of the conserved, phosphorylatable aspartate residue in the receiver domain is required for DigR activity. Consistent with the defect in type IV pili dependent motility, a digR mutant is slightly reduced in type IV pili biosynthesis and the composition of the extracellular matrix fibrils is abnormal with an increased content of polysaccharides and decreased accumulation of the FibA metalloprotease. Using genome-wide transcriptional profiling, 118 genes were identified that are directly or indirectly regulated by DigR. These genes only include two genes, agmQ and cheY4, previously implicated in A- and type IV pili dependent motility, respectively. In silico analyses showed that 36% of the differentially expressed genes are likely to encode exported proteins. Moreover, four genes encoding homologs of ECF sigma factors, which typically control aspects of cell envelope homeostasis, are differentially expressed in a digR mutant. We suggest that the DigR response regulator has an important function in cell envelope homeostasis and that the motility defects in a digR mutant are instigated by the abnormal cell envelope and abnormal expression of agmQ and cheY4. Keywords: vegetative analysis (delta)DigR / DK1622 (wt)
2007-11-02 | GSE9501 | GEO
Project description:PilY1 is a component of the type IV pili, an important virulence factor for P. aeruginosa. PilY1 has demonstrated signaling function and we wanteed to define the signal transduction pathways tht PilY1 controls. We found that PilY1 controls both cAMP and c-di-GMP. We determined the mechanism for how Pily1 influences cAMP production in P. aeruginosa.
2024-12-01 | GSE278651 | GEO
Project description:Functional characterization of a plasmid-encoded type IV-A3 CRISPR-Cas system
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