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Transcriptomic analysis reveals a novel LysR regulator's role in Burkholderia pseudomallei host-pathogen interaction

ABSTRACT: 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.

ORGANISM(S): Burkholderia pseudomallei

PROVIDER: GSE302379 | GEO | 2025/12/31

REPOSITORIES: GEO

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Project description:Burkholderia pseudomallei is a soil-dwelling bacterium which has to survive not only under harsh environmental conditions, but also within various hosts where it can cause the infectious disease melioidosis. The ability, to quickly adapt to these different conditions, is based on its huge genome which encodes for complex regulatory networks. Among them are more than 60 genes belonging to the group of LysR-type transcriptional regulators (LTTRs). Here we analyzed a B. pseudomallei mutant harboring a transposon in the gene BPSL0117 annotated as a LTTR, which we named gvmR (globally acting virulence and metabolism regulator). The gvmR mutant displayed a growth defect in minimal medium and macrophages in comparison with the wild type. Moreover, inactivation of GvmR rendered B. pseudomallei avirulent in mice indicating a critical role of GvmR in infection. These defects of the mutant were rescued by ectopic expression of gvmR. To identify genes whose expression is modulated by GvmR, global transcriptome analysis of the B. pseudomallei wild type and gvmR mutant was performed using whole genome tiling microarrays. Transcript levels of 190 and 142 genes were found to be up- and downregulated in the gvmR mutant relative to the wild type. Among the most downregulated genes in the gvmR mutant were important virulence factor genes (T3SS3, T6SS1 and T6SS2), which might provide an explanation for the virulence defect of the gvmR mutant. In addition, expression of genes related to amino acid synthesis, glyoxylate shunt, iron-sulfur cluster assambly and syrbactin metabolism (secondary metabolite) was decreased in the mutant. Furthermore, inactivation of GvmR increased expression of genes involved in pyruvate metabolism, ATP synthesis, malleobactin and porin genes. Quantitative RT-PCR verified the differential expression of 27 selected genes. In summary, our data show that GvmR acts as an activating and repressing global regulator that is required to coordinate expression of a diverse set of metabolic and virulence genes for survival in the animal host and under nutrient limitation.

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Transcriptomic analysis reveals a novel LysR regulator's role in Burkholderia pseudomallei host-pathogen interaction

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