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Mycobacterium tuberculosis tRNA induces IL-12p70 via synergistic activation of pattern recognition receptors within a cell network.

ABSTRACT: Mycobacterium tuberculosis tRNA induces IL-12p70 via synergistic activation of pattern recognition receptors within a cell network.

PROVIDER: PRJNA433477 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR6686870.fastq.gz	Fastqsanger.gz
	SRR6686880.fastq.gz	Fastqsanger.gz
	SRR6686890.fastq.gz	Fastqsanger.gz
	SRR6686871.fastq.gz	Fastqsanger.gz
	SRR6686881.fastq.gz	Fastqsanger.gz

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Mycobacterium tuberculosis tRNA induces IL-12p70 via synergistic activation of pattern recognition receptors within a cell network.

Project description:Analysis of gene expression profiles identified a mycobacterial tRNA-induced innate immune network resulting in the robust production of IL-12p70, a cytokine required to instruct an adaptive Th1 response for host defense against intracellular bacteria.

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Context-dependent compensation among phosphatidylserine-recognition receptors

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Physiological role of 2’-O-methylation within prokaryotic and eukaryotic tRNA for innate immune activation via endosomal Toll-like receptors

Project description:Bacterial RNA has emerged as important activator of innate immune responses by stimulating the endosomal Toll-like receptors TLR7 and TLR8 in humans. Guanosine 2’-O-methylation at position 18 (Gm18) in bacterial tRNA was shown to antagonize tRNA induced TLR7/8 activation, giving rise to a potential role of this modification as immune escape mechanism. This modification also occurs in eukaryotic tRNA, yet a physiological immune function remains to be established. We therefore set out to investigate the physiological role of Gm18 in prokaryotic and eukaryotic microorganisms by using mutants deficient in the respective 2’-O-methyltransferase. In E. coli, lack of 2’-O-methyltransferase trmH enhanced immune stimulatory properties of both tRNA and whole cellular RNA. Yet, when using living microorganisms, trmH mutants did not differ from their wildtype counterparts in terms of immunostimulation although gene expression profiling demonstrated the induction of a TLR8/RNA dependent gene signature by E. coli in principle. In summary, the results demonstrate that Gm18 is a global immune inhibitory RNA modification across the kingdoms and contributes to RNA recognition by innate immune cells.

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A Structural Motif for mRNA Target Recognition by a tRNA Pseudouridine Synthase

Project description:We used an in vitro pool based strategy to probe the sequence and structural requirements for mRNA target recognition by the yeast tRNA pseudouridine synthase Pus1.

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Peptidoglycan metabolite photoaffinity reporters reveal direct binding to intracellular pattern recognition receptors and Arf GTPases

Project description:Extensive structure-activity studies of iE-DAP and MDP have demonstrated their selective activation of NOD1- and NOD2-expressing cells, respectively. Nonetheless, the direct binding of iE-DAP and MDP to NOD1 and NOD2, respectively, as well as other proteins in mammalian cells has not been systematically investigated. To address the direct interaction of iE-DAP and MDP with NOD1 and NOD2 in living cells, we synthesized a series of peptidoglycan metabolite photoaffinity chemical reporters containing a diazirine for photocrosslinking in cells and an alkyne tag for bioorthogonal detection of covalently-labeled proteins. The analysis of these peptidoglycan metabolite photoaffinity reporters demonstrate that iE-DAP and MDP can directly bind to NOD1 and NOD2 in mammalian cells. The crosslinking of these peptidoglycan metabolite photoaffinity reporters only occur with active variants of NOD1 and NOD2. Beyond direct binding to NOD2, the active MDP photoaffinity reporter selectively crosslinked Arf GTPases in NOD2-expressing cells and did so most prominently with GTP-bound Arf6, revealing the Arf proteins as novel direct pattern recognition receptors. Additional labeling and co-precipitation experiments suggest that MDP induces a complex between NOD2 and GTP-Arf6. This study highlights the utility of peptidoglycan metabolite photoaffinity reporters for characterizing their direct binding to intracellular pattern recognition receptors and identifying unpredicted cellular cofactors. The applications of these microbial-specific metabolite reporters should enable the discovery of other receptors, transporters and regulatory enzymes in host cells and microbes.

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Mycobacterium tuberculosis Transfer RNA Induces IL-12p70 via Synergistic Activation of Pattern Recognition Receptors within a Cell Network.

Project description:Upon recognition of a microbial pathogen, the innate and adaptive immune systems are linked to generate a cell-mediated immune response against the foreign invader. The culture filtrate of Mycobacterium tuberculosis contains ligands, such as M. tuberculosis tRNA, that activate the innate immune response and secreted Ags recognized by T cells to drive adaptive immune responses. In this study, bioinformatics analysis of gene-expression profiles derived from human PBMCs treated with distinct microbial ligands identified a mycobacterial tRNA-induced innate immune network resulting in the robust production of IL-12p70, a cytokine required to instruct an adaptive Th1 response for host defense against intracellular bacteria. As validated by functional studies, this pathway contained a feed-forward loop, whereby the early production of IL-18, type I IFNs, and IL-12p70 primed NK cells to respond to IL-18 and produce IFN-γ, enhancing further production of IL-12p70. Mechanistically, tRNA activates TLR3 and TLR8, and this synergistic induction of IL-12p70 was recapitulated by the addition of a specific TLR8 agonist with a TLR3 ligand to PBMCs. These data indicate that M. tuberculosis tRNA activates a gene network involving the integration of multiple innate signals, including types I and II IFNs, as well as distinct cell types to induce IL-12p70.

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Project description:Plants deploy pattern recognition receptors to detect microbe- and damage-associated molecular patterns. Arabidopsis thaliana receptor-like protein RLP30 contributes to innate immunity to the necrotrophic fungus Sclerotinia sclerotiorum by recognizing SCLEROTINIA CULTURE FILTRATE ELICITOR 1 (SCFE1). Here we show that the S. sclerotiorum small cysteine-rich protein SCP1 accounts for elicitor activity of SCFE1. RLP30 recognizes SCP1 and its homologs from divergent fungi and oomycetes, as well as an SCP1-unrelated and conserved pattern from bacterial Pseudomonads. Stable expression of RLP30 in Nicotiana tabacum confers enhanced immunity to bacterial, fungal, and oomycete pathogens. Unlike Arabidopsis, which requires intact SCP1 for RLP30-mediated immunity, other Brassicaceae and Solanaceae respond to smaller immunogenic SCP1 epitopes. We conclude that Arabidopsis RLP30 recognizes immunogenic patterns from three microbial kingdoms and that mechanistically different SCP1 perception has evolved in other plant species, likely as a result of convergent evolution.

2023-06-14 | PXD036013 | Pride

Gene expression study reveals compromised Pattern Recognition Receptors and Interferon Signaling in fullterm Low birth Weight newborns

Project description:Low Birth Weight (LBW) newborns (with weight less than 2500g) suffer a higher frequency and severity of microbial infection than normal birth weight (NBW) newborns. Morbidity and mortality of LBW infants due to infectious diseases are known to be high and it has been associated with compromised immune functions, however, the complete understanding of the cause is lacking. We, therefore, conducted a gene expression study to identify all the defective pathways and functions in the LBW newborns using DNA microarray. RNA were isolated from the umbilical cord blood of the NBW and LBW newborns and processed for chip hybridisation. Our results suggest down-regulation of genes participating in several canonical pathways vital in the defense response against microbes and perpetuation of immune responses. Pattern recognition receptors (PRRs) and Interferon signaling appear to be most significantly impacted pathways. The information generated from this study may help in depth understanding of the cause of higher frequency of infections in LBW and thus, in turn help devise better therapeutic interventions. Total 12 subjects were included in the study. 4 NBW newborns umblical cord blood samples served as control and 8 LBW newborns umblical cord blood samples were treated as the experimental samples for the study.

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