College of Biology and Food, Shangqiu Normal University, Shangqiu 476000, Henan Provence, China; College of Medical Technology, Shangqiu Medical College, Shangqiu 476005, Henan Provence, China.
College of Biology and Food, Shangqiu Normal University, Shangqiu 476000, Henan Provence, China.
J Proteomics. 2024 Oct 30;309:105297. doi: 10.1016/j.jprot.2024.105297. Epub 2024 Aug 28.
Brucella spp. is an intracellular bacterium that uses its transcriptional regulator DeoR1 to promote intracellular transport and survival, but the molecular mechanism remains unknown. To analyze the role of DeoR1 in the virulence of B. abortus and the genes regulated by DeoR1, we created a A19ΔdeoR1 mutant of B. abortus A19 (A19). Virulence assay was performed using a murine macrophage cell line (RAW264.7) and mice. We observed that A19ΔdeoR1 mutant is attenuated in RAW264.7 cells and mice. We performed RNA-seq whole transcriptome analysis of A19ΔdeoR1 and A19 from infected RAW264.7 cells. A total of 135 differentially expressed genes were identified, including 100 up-regulated and 35 down-regulated genes. These differentially expressed genes were involved in amino acid synthesis and metabolism, energy production and conversion, stress proteins, chaperonin, hypothetical proteins and protein of unknown function, cell wall/membrane/envelope, intracellular transporting and secretion, and transcriptional regulator. Interestingly, genes involved in the intracellular trafficking and secretion were significantly down-regulated in A19ΔdeoR1. Furthermore, selected RNA-seq results were experimentally confirmed by qRT-PCR. Overall, these results deciphered differential phenomena associated with virulence in A19ΔdeoR1 and A19 from infected RAW264.7 cells, which provided important information for understanding the detailed role of DeoR1 in Brucella pathogenesis. SIGNIFICANCE: Transcriptional regulators are predominant bacterial signal transduction factors. The pathogenicity of Brucella is due to its ability to regulate the expression of virulence related genes. Transcriptional regulators are designed to regulate gene expression and enact an appropriate adaptive physiological response. Here, a total of 135 differentially expressed genes were identified in transcriptional regulator deoR1 mutant.
布鲁氏菌属是一种细胞内细菌，它利用其转录调节因子 DeoR1 促进细胞内运输和存活，但分子机制尚不清楚。为了分析 DeoR1 在流产布鲁氏菌 A19 (A19) 毒力和 DeoR1 调节的基因中的作用，我们构建了 A19 的 A19ΔdeoR1 突变体。使用鼠巨噬细胞系（RAW264.7）和小鼠进行了毒力测定。我们观察到 A19ΔdeoR1 突变体在 RAW264.7 细胞和小鼠中减毒。我们对感染 RAW264.7 细胞的 A19ΔdeoR1 和 A19 进行了 RNA-seq 全转录组分析。共鉴定出 135 个差异表达基因，包括 100 个上调和 35 个下调基因。这些差异表达基因参与氨基酸合成和代谢、能量产生和转化、应激蛋白、伴侣蛋白、假定蛋白和未知功能蛋白、细胞壁/膜/包膜、细胞内运输和分泌以及转录调节因子。有趣的是，A19ΔdeoR1 中参与细胞内运输和分泌的基因显著下调。此外，通过 qRT-PCR 实验验证了 RNA-seq 结果。总的来说，这些结果揭示了 A19ΔdeoR1 和感染 RAW264.7 细胞的 A19 之间与毒力相关的差异现象，为理解 DeoR1 在布鲁氏菌发病机制中的详细作用提供了重要信息。意义：转录调节因子是主要的细菌信号转导因子。布鲁氏菌的致病性归因于其调节与毒力相关基因表达的能力。转录调节因子旨在调节基因表达并引发适当的适应性生理反应。在这里，在转录调节因子 deoR1 突变体中鉴定出总共 135 个差异表达基因。
