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首页> 外文期刊>Fish & Shellfish Immunology >Integrated dual RNA-seq and dual iTRAQ of infected tissue reveals the functions of a diguanylate cyclase gene of Pseudomonas plecoglossicida in host-pathogen interactions with Epinephelus coioides
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Integrated dual RNA-seq and dual iTRAQ of infected tissue reveals the functions of a diguanylate cyclase gene of Pseudomonas plecoglossicida in host-pathogen interactions with Epinephelus coioides

机译：综合的双RNA-SEQ和受感染组织的双ITRAQ揭示了与EPINEPHELUS COIIAIDE的宿主病原体相互作用的Pseudomonaspocoglossica的Diguantylys Cycoglossica的功能

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摘要

The interactions between host and pathogen is exceedingly complex, which involves alterations at multiple molecular layers. However, research to simultaneously monitor the alterations of transcriptome and proteome between a bacterial pathogen and aquatic animal host through integrated dual RNA-seq and dual iTRAQ of tissue during infection is currently lacking. The important role of a diguanylate cyclase gene (L321_RS15240) in pathogenicity of Pseudomonas plecoglossicida against Epinephelus coioides was suggested by previous dual RNA-seq of our lab. Then L321_RS15240-RNAi strains of P. plecoglossicida were constructed with pCM130/tac, and the mutant with the best silencing effect was selected for follow-up study. The RNAi of L321_RS15240 resulted in a significant decrease in bacterial virulence of P. plecoglossicida. The E. coioides spleens infected by wild type strain or L321_RS15240-RNAi strain of P. plecoglossicida were subjected to dual RNA-seq and dual iTRAQ, respectively. The results showed that: RNAi of L321_RS15240 led to 1)alterations of host transcriptome associated with complement and coagulation cascades, ribosome, arginine and proline metabolism, and oxidative phosphorylation; 2)high expression of host proteins which related to phagosome and metabolism responses (metabolism of glutathione, amino sugar and nucleotide sugar); 3)the highly differentially expression of host lncRNAs and miRNAs. The differentially expressed proteins and mRNAs of pathogen were different after infection, but the functions of these proteins and mRNAs were mainly related to metabolism and virulence. This study provides a new insight to comprehensively understand the gene functions of pathogens and hosts at multiple molecular layers during in vivo infection.
机译：宿主和病原体之间的相互作用非常复杂，这涉及多种分子层的改变。然而，通过在感染期间通过集成的双RNA-SEQ和组织的组织进行细菌病原体和水生动物宿主在细菌病原体和水生动物宿主之间进行调查。通过我们实验室的先前双RNA-SEQ提出了DiguantyLys Cycoglossica致肾上腺素CoioIDA的致肾上腺素Coio的重要作用。然后用PCM130 / TAC构建L321_RS15240-RNAI菌株，并选择具有最佳沉默效果的突变体进行后续研究。 L321_RS15240的RNAi导致P.Plecoglossicida的细菌毒力显着降低。受野生型菌株或L321_RS15240-RNAI菌株的脾脏的E. CoIoides分别对P.Plecoglossicida进行双RNA-SEQ和双Itraq。结果表明：L321_RS15240的RNAi LED为1）宿主转录组的改变与补体和凝固级联，核糖体，精氨酸和脯氨酸代谢和氧化磷酸化相关; 2）与吞噬蛋白酶和代谢反应有关的宿主蛋白的高表达（谷胱甘肽，氨基糖和核苷酸糖的代谢）; 3）宿主LNCRNA和miRNA的高差异表达。感染后，病原体的差异表达蛋白质和mRNA不同，但这些蛋白质和MRNA的功能主要与代谢和毒力有关。本研究提供了新的洞察，以全面了解病原体和宿主在体内感染期间多种分子层的基因功能。

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