College of Animal & Veterinary Sciences, Key Laboratory of Animal Medicine of Sichuan Province, Southwest Minzu University, Chengdu, China.
The State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Hubei Hongshan Laboratory, Huazhong Agricultural University, Wuhan, China.
mSystems. 2024 Apr 16;9(4):e0089123. doi: 10.1128/msystems.00891-23. Epub 2024 Mar 5.
species are able to produce and release secreted proteins, such as toxins, adhesins, and virulence-related enzymes, involved in bacteria adhesion, invasion, and immune evasion between the pathogen and host. Here, we investigated a novel secreted protein, MbovP0725, from encoding a putative haloacid dehalogenase (HAD) hydrolase function of a key serine/threonine phosphatase depending on Mg for the dephosphorylation of its substrate and it was most active at pH 8 to 9 and temperatures around 40°C. A transposon insertion mutant strain of HB0801 that lacked the protein MbovP0725 induced a stronger inflammatory response but with a partial reduction of adhesion ability. Using transcriptome sequencing and quantitative reverse transcription polymerase chain reaction analysis, we found that the mutant was upregulated by the mRNA expression of genes from the glycolysis pathway, while downregulated by the genes enriched in ABC transporters and acetate kinase-phosphate acetyltransferase pathway. Untargeted metabolomics showed that the disruption of the gene caused the accumulation of 9-hydroxyoctadecadienoic acids and the consumption of cytidine 5'-monophosphate, uridine monophosphate, and adenosine monophosphate. Both the exogenous and endogenous MbvoP0725 protein created by purification and transfection inhibited lipopolysaccharide (LPS)-induced IL-1β, IL-6, and TNF-α mRNA production and could also attenuate the activation of MAPK-associated pathways after LPS treatment. A pull-down assay identified MAPK p38 and ERK as potential substrates for MbovP0725. These findings define metabolism- and virulence-related roles for a HAD family phosphatase and reveal its ability to inhibit the host pro-inflammatory response.
() infection is characterized by chronic pneumonia, otitis, arthritis, and mastitis, among others, and tends to involve the suppression of the immune response via multiple strategies to avoid host cell immune clearance. This study found that MbovP0725, a haloacid dehalogenase (HAD) family phosphatase secreted by , had the ability to inhibit the host pro-inflammatory response induced by lipopolysaccharide. Transcriptomic and metabolomic analyses were used to identify MbovP0725 as an important phosphatase involved in glycolysis and nucleotide metabolism. The transposon mutant strain T8.66 lacking MbovP0725 induced a higher inflammatory response and exhibited weaker adhesion to host cells. Additionally, T8.66 attenuated the phosphorylation of MAPK P38 and ERK and interacted with the two targets. These results suggested that MbovP0725 had the virulence- and metabolism-related role of a HAD family phosphatase, performing an anti-inflammatory response during infection.
物种能够产生和释放分泌蛋白，如毒素、黏附素和与毒力相关的酶，这些蛋白参与病原体与宿主之间的细菌黏附、入侵和免疫逃避。在这里，我们研究了一种来自 的新型分泌蛋白 MbovP0725，它编码一种假定的 haloacid dehalogenase (HAD) 水解酶，该酶依赖于 Mg 发挥关键丝氨酸/苏氨酸磷酸酶的功能，使底物去磷酸化，其最适 pH 值为 8 到 9，最适温度约为 40°C。HB0801 的转座子插入突变株缺乏蛋白 MbovP0725，它诱导更强的炎症反应，但黏附能力部分降低。通过转录组测序和定量逆转录聚合酶链反应分析，我们发现突变体中与糖酵解途径相关的基因的 mRNA 表达上调，而与 ABC 转运蛋白和乙酸激酶-磷酸乙酰转移酶途径相关的基因下调。非靶向代谢组学表明， 基因的破坏导致 9-羟基十八碳二烯酸的积累和胞苷 5'-单磷酸、尿苷单磷酸和腺苷单磷酸的消耗。纯化和转染产生的外源性和内源性 MbvoP0725 蛋白均能抑制脂多糖 (LPS) 诱导的 IL-1β、IL-6 和 TNF-αmRNA 的产生，并能减轻 LPS 处理后 MAPK 相关途径的激活。下拉实验鉴定 MAPK p38 和 ERK 为 MbovP0725 的潜在底物。这些发现定义了 HAD 家族磷酸酶与代谢和毒力相关的作用，并揭示了其抑制宿主促炎反应的能力。
()感染的特征是慢性肺炎、中耳炎、关节炎和乳腺炎等，并且往往通过多种策略来抑制免疫反应，以避免宿主细胞的免疫清除。本研究发现，由 分泌的 haloacid dehalogenase (HAD) 家族磷酸酶 MbovP0725 具有抑制脂多糖诱导的宿主促炎反应的能力。使用转录组和代谢组学分析鉴定 MbovP0725 是参与糖酵解和核苷酸代谢的重要磷酸酶。缺失 MbovP0725 的 转座子突变株 T8.66 诱导更高的炎症反应，对宿主细胞的黏附能力减弱。此外，T8.66 减弱了 MAPK P38 和 ERK 的磷酸化，并与这两个靶点相互作用。这些结果表明，MbovP0725 具有 HAD 家族磷酸酶的毒力和代谢相关作用，在 感染过程中发挥抗炎反应。
