胆固醇依赖性 Nsp5-内体向溶酶体的共同运输通过激活自噬促进猪繁殖与呼吸综合征病毒的复制

IF 2.4 2区农林科学 Q3 MICROBIOLOGY

Veterinary microbiology Pub Date : 2025-04-08 DOI:10.1016/j.vetmic.2025.110507

Haoxin Dong , Qiming Pei , Jiahui Ren , Yaci Zhang , Xuedan Wei , Aijuan Shen , Yunshuo Lu , Ziheng Zhang , Yongkun Du , Guoqing Zhuang , Angke Zhang , Hong Duan

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引用次数: 0

摘要

我们之前的研究表明，在病毒感染的早期，细胞内的内体囊泡参与了PRRS病毒粒子的运输，并且内体囊泡中的胆固醇滞留通过阻断prrsv -内体囊泡膜融合来干扰病毒的复制。然而，内体囊泡是否与PRRSV蛋白运输相关以及胆固醇在这一过程中的作用尚不清楚。在这项研究中，我们试图阐明胆固醇在内体囊泡介导的病毒蛋白运输中的机制。结果表明，内体囊泡参与了PRRSV Nsp5蛋白的转运。在内质网（ER）和高尔基体中合成后，Nsp5被运输到早期核内体（EEs），而不是内吞循环室（ERCs），然后运输到晚期核内体（LEs），最终到达溶酶体室，而胆固醇通量或LEs功能的破坏导致Nsp5无法到达溶酶体，在那里Nsp5激活细胞自噬以促进PRRSV复制。分子对接预测表明，胆固醇可以与Nsp5的74丙氨酸和78天冬酰胺形成两个氢键。在上述结合位点发生突变后，PRRSV的复制效率降低。随后，研究人员探索了胆固醇在PRRSV复制中的作用。阻断胆固醇通量可显著抑制PRRSV复制。单病毒感染周期分析表明，胆固醇通量紊乱不影响病毒吸附，但可以抑制病毒进入宿主细胞，阻断ees - les -溶酶体介导的病毒粒子运输，导致病毒粒子滞留在内体腔室中。目前的研究表明，胆固醇和内体囊泡协同参与Nsp5转运，促进PRRSV复制，这可能为开发针对胆固醇代谢途径的新型抗病毒药物或改进商业疫苗提供新的见解。

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Cholesterol-dependent Nsp5-endosomes co-trafficking to lysosomes facilitates porcine reproductive and respiratory syndrome virus replication by activating autophagy

Our previous studies showed that intracellular endosomal vesicles participated in PRRS virions trafficking in the early stage of viral infection, and cholesterol retention in endosomal vesicles disturbed viral replication via blocking PRRSV-endosomal vesicles membrane fusion. However, whether endosomal vesicles were associated with PRRSV protein(s) trafficking and the role of cholesterol in this process was still unclarity. In this study, we sought to elucidate the mechanism of cholesterol in endosomal vesicles-mediated viral protein transportation. The results showed that endosomal vesicles participated in trafficking of PRRSV Nsp5 protein. After being synthesized in endoplasmic reticulum (ER) and Golgi apparatus, Nsp5 was trafficked to early endosomes (EEs), but not endocytic recycling compartments (ERCs), then to late endosomes (LEs), and eventually reached lysosomal compartments, whereas disruption of cholesterol flux or LEs function led to the inability of Nsp5 arriving at lysosomes, where Nsp5 activated cellular autophagy to promote PRRSV replication. Molecular docking predictions revealed that cholesterol could form two hydrogen bonds with 74 alanine and 78 asparagine of Nsp5. After mutating the aforementioned binding sites, the replication efficiency of PRRSV decreased. Subsequently, the role of cholesterol in PRRSV replication was explored. Blocking of cholesterol flux significantly inhibited PRRSV replication. Single virus infection cycle analysis showed that cholesterol flux disorder did not affect virus adsorption, but could inhibit virus entry into host cells and block EEs-LEs-lysosomes mediated trafficking of virions, leading to virions retention in endosomal compartments. The present studies suggest that cholesterol and endosomal vesicles synergistically participate in Nsp5 trafficking to promote PRRSV replication, which may provide new insights for the development of novel antiviral drugs targeting cholesterol metabolism pathways or the improvement of commercial vaccines.

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来源期刊

Veterinary microbiology 农林科学-兽医学

CiteScore

5.90

自引率

6.10%

发文量

221

审稿时长

52 days

期刊介绍： Veterinary Microbiology is concerned with microbial (bacterial, fungal, viral) diseases of domesticated vertebrate animals (livestock, companion animals, fur-bearing animals, game, poultry, fish) that supply food, other useful products or companionship. In addition, Microbial diseases of wild animals living in captivity, or as members of the feral fauna will also be considered if the infections are of interest because of their interrelation with humans (zoonoses) and/or domestic animals. Studies of antimicrobial resistance are also included, provided that the results represent a substantial advance in knowledge. Authors are strongly encouraged to read - prior to submission - the Editorials (''Scope or cope'' and ''Scope or cope II'') published previously in the journal. The Editors reserve the right to suggest submission to another journal for those papers which they feel would be more appropriate for consideration by that journal. Original research papers of high quality and novelty on aspects of control, host response, molecular biology, pathogenesis, prevention, and treatment of microbial diseases of animals are published. Papers dealing primarily with immunology, epidemiology, molecular biology and antiviral or microbial agents will only be considered if they demonstrate a clear impact on a disease. Papers focusing solely on diagnostic techniques (such as another PCR protocol or ELISA) will not be published - focus should be on a microorganism and not on a particular technique. Papers only reporting microbial sequences, transcriptomics data, or proteomics data will not be considered unless the results represent a substantial advance in knowledge. Drug trial papers will be considered if they have general application or significance. Papers on the identification of microorganisms will also be considered, but detailed taxonomic studies do not fall within the scope of the journal. Case reports will not be published, unless they have general application or contain novel aspects. Papers of geographically limited interest, which repeat what had been established elsewhere will not be considered. The readership of the journal is global.