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ABSTRACT: Plaque-forming-defective Burkholderia pseudomallei

PROVIDER: PRJNA635029 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR11848389.fastq.gz	Fastqsanger.gz
	SRR11848390.fastq.gz	Fastqsanger.gz

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Toxicogenomic Effect of Burkholderia pseudomallei in a Human Macrophage Cell (THP-1) Model of Acute Melioidosis Using Whole-genome Microarray

Project description:Gene expression profiles of human cell (THP-1) lines exposed to a novel Daboiatoxin (DbTx) isolated from Daboia russelli russelli, and specific cytokines and inflammatory pathways involved in acute infection caused by Burkholderia pseudomallei. Keywords: Melioidosis, Burkholderia pseudomallei, Daboiatoxin, Cytokines, Inflammation.

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Project description:Burkholderia pseudomallei is a Gram-negative pathogen responsible for melioidosis, a life-threatening disease endemic to Southeast Asia. LysR-type transcriptional regulators (LTTRs) are known key regulators of bacterial pathogenesis and metabolism, yet many remain uncharacterized. This study investigates the function of UKMD286_5923, a novel LTTR, in B. pseudomallei UKMD286. We constructed a deletion mutant and performed transcriptomic analysis via RNA sequencing. This analysis identified 67 differentially expressed genes, with 45 genes upregulated and 22 genes downregulated in the mutant compared to the wild-type. Functional enrichment analysis of these genes highlighted significant roles in metabolism, transport and secretion system. To further characterize the phenotypic impact of the gene deletion, we conducted biofilm formation and plaque assays. Biofilm formation assays showed increased biofilm production in the mutant, suggesting a regulatory role in bacterial adhesion. Plaque assays revealed reduced plaque formation in the mutant, indicating impaired host cell invasion. These findings collectively suggest that UKMD286_5923 influences genes essential for bacterial survival and host-pathogen interaction, including components of the Type III secretion system. Understanding the function of this regulator enhances our knowledge of B. pseudomallei pathogenesis and may contribute to future diagnostic and treatment strategies for melioidosis.

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Project description:We report the application of single-molecule-based sequencing technology for high-throughput profiling of DNA methylations in Burkholderia pseudomallei.

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Project description:Melioidosis, caused by the Gram-negative bacterium Burkholderia pseudomallei, is a disease endemic to South-East Asia and Northern Australia. Clinical presentation is highly variable, ranging from asymptomatic to fatal septicaemia, and thus the outcome of infection can depend on the host immune responses. The aim of this study was to characterise the macrophage immune response to B. pseudomallei in the presence of novel inhibitors targeting the virulence factor, Macrophage Infectivity Potentiator (Mip) protein. To do this. murine macrophage J774A.1 cells were infected with B. pseudomallei K96243 in the presence and absence of two small-molecule inhibitors designed to target the Mip protein. Global transcriptional profiling of macrophages infected with B. pseudomallei was analysed by RNA-Seq four hours post-infection. In the presence of Mip inhibitors, we found a significant reduction in the expression of pro-inflammatory cytokines highlighting the potential to utilize Mip inhibitors to dampen potentially harmful pro-inflammatory responses resulting from B. pseudomallei infection in macrophages. We then performed gene expression profiling analysis using data obtained from RNA-seq of J774A.1 macrophages infected with Burkholderia pseudomallei in the presence of two Mip inhibitors or vehicle control 4 hours post-infection

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