Transcriptional regulator GntR controls diverse physiological functions necessary for Brucella survival. In the intracellular pathogen Brucella, GntR has been shown to regulate the expression of genes related to virulence. However, the precise determination of GntR direct targets has so far proved elusive. Therefore, we performed chromatin immunoprecipitation of GntR10 followed by next‐generation sequencing (ChIP‐seq). We selected target gene BAB1_1163 directly regulated by GntR10 and created the mutant (2308Δ1163) from virulent Brucella abortus 2308 (S2308). 2308Δ1163 strain survival capability in murine macrophages (RAW 264.7) was detected and the levels of tumor necrosis factor‐α (TNF‐α) and interleukin‐1β (IL‐1β) were also measured. We detected 88 intergenic ChIP‐seq peaks of GntR10 binding distributed across the Brucella genome and determined a markedly asymmetric binding consensus motif with 12 bp length. 2308Δ1163 showed reduced survival capability in RAW 264.7. After the macrophages were infected with 2308Δ1163, the levels of TNF‐α and IL‐1β were decreased and were significantly lower than that for the S2308‐infected group, indicating that the 2308Δ1163 mutant could inhibit the secretion of inflammatory cytokines. Taken together, the research has recorded valuable data about GntR10 and provided new insights into the functionality of GntR10.

中文翻译：

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布鲁氏菌的 ChIP-seq 分析揭示了 GntR 的转录调控

转录调节因子 GntR 控制布鲁氏菌存活所需的多种生理功能。在胞内病原体布鲁氏菌中，GntR 已被证明可以调节与毒力相关的基因的表达。然而，迄今为止，GntR 直接目标的精确确定难以实现。因此，我们对 GntR10 进行染色质免疫沉淀，然后进行二代测序（ChIP-seq）。我们选择了由 GntR10 直接调控的靶基因 BAB1_1163，并从有毒的流产布鲁氏菌 2308 (S2308) 中产生了突变体 (2308Δ1163)。检测了小鼠巨噬细胞 (RAW 264.7) 中 2308Δ1163 菌株的存活能力，并测量了肿瘤坏死因子-α (TNF-α) 和白细胞介素-1β (IL-1β) 的水平。我们检测到分布在布鲁氏菌基因组中的 88 个 GntR10 结合的基因间 ChIP-seq 峰，并确定了一个长度为 12 bp 的明显不对称的结合共有基序。2308Δ1163 在 RAW 264.7 中的生存能力下降。巨噬细胞感染2308Δ1163后，TNF-α和IL-1β水平降低，明显低于S2308感染组，表明2308Δ1163突变体可以抑制炎性细胞因子的分泌。综上所述，该研究记录了有关 GntR10 的宝贵数据，并提供了对 GntR10 功能的新见解。TNF-α 和 IL-1β 水平降低且显着低于 S2308 感染组，表明 2308Δ1163 突变体可以抑制炎性细胞因子的分泌。综上所述，该研究记录了有关 GntR10 的宝贵数据，并提供了对 GntR10 功能的新见解。TNF-α 和 IL-1β 水平降低且显着低于 S2308 感染组，表明 2308Δ1163 突变体可以抑制炎性细胞因子的分泌。综上所述，该研究记录了有关 GntR10 的宝贵数据，并提供了对 GntR10 功能的新见解。