Genome-Wide Analysis Reveals that PhoP Regulates Pathogenicity in Riemerella anatipestifer.
Genome-Wide Analysis Reveals that PhoP Regulates Pathogenicity in Riemerella anatipestifer.
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全基因组分析揭示 PhoP 调节鸭疫里默氏菌的致病性
DOI：
10.1128/spectrum.01883-22
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发表时间：
2022-10-26
影响因子：
3.7
通讯作者：
Li, Zili
中科院分区：
文献类型：
--
作者：
Zhang, Yang;Wang, Ying;Zhang, Yanhao;Jia, Xiangchao;Li, Chenxi;Zhou, Zutao;Hu, Sishun;Li, Zili
鸭疫里默氏杆菌（Riemerella anatipestifer）是一种重要的致病菌，可引起鸭、鸡、鹅等禽类的急性败血症和传染性多浆膜炎，具有很高的致死率。以前，我们的特点是由phoPR编码的双组分系统，并发现R。phoPR基因缺失后，鸭瘟病菌对雏鸭的致病性几乎完全丧失。鸭传染性浆膜炎，又称鸭疫里默氏杆菌病，主要感染家鸭、鹅、火鸡和野鸟。然而，其致病性的调控机制仍不清楚。PhoPR双组分系统（TCS）是我们前期研究中首次在革兰氏阴性菌中发现的，并被证实与致病性和基因表达有关。本研究采用DNA亲和纯化测序（DAP-seq）技术，进一步探讨PhoPR基因在致病性调控中的作用。鸭疫在583个受PhoP直接调控的候选靶基因上游发现了一个保守基序。为了进一步确认受PhoR和PhoP调控的基因，构建了单基因缺失菌株。使用新一代RNA测序的转录组分析结果显示，ΔphoP菌株和野生型（WT）之间存在136个差异表达基因（DEG），ΔphoR菌株和WT之间存在183个差异表达基因（DEG）。结合转录组分析和DAP-seq进一步鉴定了PhoP的候选靶基因，揭示了PhoP的主要直接调控子位于细胞膜上，并且PhoP参与了耐氧性的调控。利用鸭体内模型，发现ΔphoP和ΔphoR突变体的致病性显著低于WT。总之，我们的研究结果提供了对PhoP的直接调节的见解，并表明phoPR对R.鸭疫该基因缺失株有望成为R.可作为表达外源抗原的理想基因工程载体菌株。重要性鸭疫里默氏杆菌是家禽业中具有高死亡率的重要病原体，其可导致鸭、鸡、鹅和其他禽类物种的急性败血症和传染性多浆膜炎。以前，我们的特点是由phoPR编码的双组分系统，并发现R。phoPR基因缺失后，鸭瘟病菌对雏鸭的致病性几乎完全丧失。PhoPR调控红曲霉毒力的机制尚不清楚。对鸭疫病菌的研究还不深入。在这项研究中，我们利用DAP-seq来探索PhoP作为全球基因组中的反应调节剂的DNA结合位点。此外，分别缺失phoP和phoR，并对相应基因缺失菌株进行转录组学分析。我们确定了一系列的PhoPR双组分系统的直接调控基因。雏鸭模型表明，PhoP和PhoR都是R.鸭疫
Riemerella anatipestifer is a significant pathogen with high mortality in the poultry industry that causes acute septicemia and infectious polyserositis in ducks, chickens, geese, and other avian species. Previously, we characterized the two-component system encoded by phoPR and found that R. anatipestifer almost completely lost its pathogenicity for ducklings when phoPR was deleted. ABSTRACT Duck infectious serositis, also known as Riemerella anatipestifer disease, infects domestic ducks, geese, and turkeys and wild birds. However, the regulatory mechanism of its pathogenicity remains unclear. The PhoPR two-component system (TCS) was first reported in Gram-negative bacteria in our previous research and was demonstrated to be involved in virulence and gene expression. Here, DNA affinity purification sequencing (DAP-seq) was applied to further explore the regulation of PhoPR in relation to pathogenicity in R. anatipestifer. A conserved motif was identified upstream of 583 candidate target genes which were directly regulated by PhoP. To further confirm the genes which are regulated by PhoR and PhoP, single-gene-deletion strains were constructed. The results of transcriptome analysis using next-generation RNA sequencing showed 136 differentially expressed genes (DEGs) between the ΔphoP strain and the wild type (WT) and 183 DEGs between the ΔphoR strain and the WT. The candidate target genes of PhoP were further identified by combining transcriptome analysis and DAP-seq, which revealed that the main direct regulons of PhoP are located on the membrane and PhoP is involved in regulating aerotolerance. Using the in vivo duck model, the pathogenicity of ΔphoP and ΔphoR mutants was found to be significantly lower than that of the WT. Together, our findings provide insight into the direct regulation of PhoP and suggest that phoPR is essential for the pathogenicity of R. anatipestifer. The gene deletion strains are expected to be candidate live vaccine strains of R. anatipestifer which can be used as ideal genetic engineering vector strains for the expression of foreign antigens. IMPORTANCE Riemerella anatipestifer is a significant pathogen with high mortality in the poultry industry that causes acute septicemia and infectious polyserositis in ducks, chickens, geese, and other avian species. Previously, we characterized the two-component system encoded by phoPR and found that R. anatipestifer almost completely lost its pathogenicity for ducklings when phoPR was deleted. However, the mechanism of PhoPR regulation of virulence in R. anatipestifer had not been deeply explored. In this study, we utilized DAP-seq to explore the DNA-binding sites of PhoP as a response regulator in the global genome. Furthermore, phoP and phoR were deleted separately, and transcriptomics analysis of the corresponding gene deletion strains was performed. We identified a series of directly regulated genes of the PhoPR two-component system. The duckling model showed that both PhoP and PhoR are essential virulence-related factors in R. anatipestifer.
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