Suppression of Virulent Porcine Epidemic Diarrhea Virus Proliferation by the PI3K/Akt/GSK-3α/β Pathway

Kong, Ning; Wu, Yongguang; Meng, Qiong; Wang, Zhongze; Zuo, Yewen; Pan, Xi; Wu, Tong; Zheng, Hao; Li, Guoxin; Shen, Yang; Yu, Hai; Zhou, En‐Min; Shan, Tongling; Tong, Guangzhi

doi:10.1371/journal.pone.0161508

Cited by 35 publications

(22 citation statements)

References 53 publications

(54 reference statements)

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“…We screened potential proteins for their ability to inhibit PEDV replication using the proteomics approach of isobaric tagging for relative and absolute quantitation (iTRAQ). Vero cells were collected and assayed after infection with PEDV (strain JS-2013) at an MOI (multiplicity of infection) of 1, as previously described [43]. We found that an IFN-inducible restriction factor, BST2, was upregulated in Vero cells during PEDV infection.…”

Section: Pedv Infection Upregulates the Expression Of Bst2 In Vero Anmentioning

confidence: 91%

“…Cells grown to 50%-60% confluence were transfected with siRNA using Lipofectamine RNAiMAX (Invitrogen, 13,778,150), according to the manufacturer's instructions. The PEDV variant strain JS-2013 was isolated and stored in our laboratory [43]. According to the requirements of different experiments, Vero cells were infected with PEDV at an MOI of 1 or 0.01, or were mock infected with DMEM.…”

Section: Antibodies Reagents and Plasmidsmentioning

confidence: 99%

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BST2 suppresses porcine epidemic diarrhea virus replication by targeting and degrading virus nucleocapsid protein with selective autophagy

et al. 2019

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Interferon-induced BST2 (bone marrow stromal cell antigen 2) inhibits viral replication by tethering enveloped virions to the cell surface to restrict viral release and by inducing the NFKB-dependent antiviral immune response. However, the mechanism by which BST2 uses the selective autophagy pathway to inhibit viral replication is poorly understood. In this study, we showed that BST2 expression was significantly increased during porcine epidemic diarrhea virus (PEDV) infection of Vero cells by IRF1 targeting its promoter. We also showed that BST2 suppressed PEDV replication by binding and degrading the PEDV-encoded nucleocapsid (N) protein. The downregulation of N protein was blocked by macroautophagy/autophagy inhibitors but not a proteasome inhibitor, implying that the N protein was degraded via the selective autophagy pathway. Both the BST2 and N protein interacted with the E3 ubiquitin ligase MARCHF8/MARCH8 and the cargo receptor CALCOCO2/NDP52, and the ubiquitination of N protein was necessary for the degradation of N mediated by the BST2-MARCHF8 axis. The knockdown of MARCHF8 or ATG5 with small interfering RNAs blocked the selective autophagy pathway, rescued the protein abundance of PEDV N in 293T cells, and prevented the inhibition of PEDV replication by BST2 in Vero cells. Together, our data demonstrate the novel mechanism of BST2-mediated virus restriction, in which BST2 recruits MARCHF8 to catalyze the ubiquitination of the PEDV N protein. The ubiquitinated N protein is then recognized by CALCOCO2/NDP52, which delivers it to autolysosome for degradation through the selective autophagy pathway.

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Section: Pedv Infection Upregulates the Expression Of Bst2 In Vero Anmentioning

confidence: 91%

Section: Antibodies Reagents and Plasmidsmentioning

confidence: 99%

BST2 suppresses porcine epidemic diarrhea virus replication by targeting and degrading virus nucleocapsid protein with selective autophagy

et al. 2019

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“…To analyze the anti-sera or antibodies specificity interacted with PEDV, Vero cells were infected with PEDV (multiplicity of infection, MOI = 1), or mock infected with the medium, and then incubated for indicated times as previously described [16]. The cells were harvested and lysed using RIPA lysis buffer (Thermo, USA) containing protease inhibitor cocktail (Bimake, USA) and phosphatase inhibitor cocktail (Bimake, USA) on ice for 5 min.…”

Section: Virus Infection and Western Blot Analysismentioning

confidence: 99%

Identification of a novel B-cell epitope in the spike protein of porcine epidemic diarrhea virus

Kong

Meng

Jiao

et al. 2020

Virol J

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Background: Porcine epidemic diarrhea virus (PEDV) infection causes an acute enteric tract infectious disease characterized by vomiting, anorexia, dehydration, weight loss and high mortality in neonatal piglets. During PEDV infection, the spike protein (S) is a major virion structural protein interacting with receptors and inducing neutralizing antibodies. However, the neutralizing B-cell epitopes within PEDV S protein have not been well studied.Methods: To accurately identify the important immunodominant region of S1, the purified truncated S1 proteins (SA, SB, SC, SD and SE) were used to immunize BALB/c mice to prepare polyclonal antibodies. The antisera titers were determined by indirect ELISA, western blot and IFA after four immunizations to find the important immunodominant region of S1, and then purified the immunodominant region of S1 protein and immunized mice to generate the special antibodies, and then used recombinant peptides to determine the B-cell epitopes of monoclonal antibodies.Results: Five antisera of recombinant proteins of the spike protein region of PEDV were generated and we found that only the polyclonal antibody against part of the S1 region (signed as SE protein, residues 666-789) could recognize the native PEDV. Purified SE protein was used to immunize BALB/c mice and generate mAb 2E10. Pepscan of the SE protein demonstrated that SE16 ( 722 SSTFNSTREL 731 ) is the minimal linear epitope required for reactivity with the mAb 2E10. Further investigation indicated that the epitope SE16 was localized on the surface of PEDV S protein in the 3D structure.Conclusions: A mAb 2E10 that is specifically bound to PEDV was generated and identified a specific linear B-cell epitope (SE16, 722 SSTFNSTREL 731 ) of the mAb. The epitope region of PEDV S1 localized in the different regions in comparison with the earlier identified epitopes. These findings enhance the understanding of the PEDV spike protein structure for vaccine design and provide a potential use for developing diagnostic methods to detect PEDV.

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“…In addition, an increasing number of studies have demonstrated that viruses encode defensive mechanisms to evade the antiviral activities of host cells (Xing et al, 2013;Yang et al, 2018). However, hosts have many mechanisms aimed at detecting and disrupting PEDV replication and spread (Cao et al, 2015;Kong et al, 2016). An improved understanding of how this pathogen interacts with the host immune system is required to enhance efforts to control PEDV spread and to develop efficacious PEDV vaccines.…”

Section: Introductionmentioning

confidence: 99%

Tumor suppressor p53 inhibits porcine epidemic diarrhea virus infection via interferon-mediated antiviral immunity

Hao

Cao

et al. 2019

Molecular Immunology

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A B S T R A C T p53 is a tumor suppressor gene that can be activated in many contexts, such as DNA damage or stressful conditions. p53 has also been shown to be important for responses to certain viral infections. Porcine epidemic diarrhea virus (PEDV) is a major enteric pathogen of the coronavirus family that causes extensive mortality among piglets. The involvement of p53 during PEDV infection has not previously been investigated. In this study, we detected p53 upregulation in response to PEDV infection. Treatment with a p53 specific activator or p53 overexpression markedly decreased viral replication, and we showed that there was more viral progeny produced in p53 knock-out cells than in p53 wild-type cells. Finally, we demonstrated that inhibition of viral infection by p53 was mediated via p53-dependent IFN signaling, leading to IFN-stimulated response element (ISRE) activation, as well as the upregulation of IFN-stimulated genes (ISGs) and IFN-β released from infected cells. These findings demonstrate that p53 suppresses PEDV infection, offering a novel therapeutic strategy for combatting this deadly disease in piglets. as a central regulator of the innate immune response (Rivas et al., https://doi.

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Suppression of Virulent Porcine Epidemic Diarrhea Virus Proliferation by the PI3K/Akt/GSK-3α/β Pathway

Cited by 35 publications

References 53 publications

BST2 suppresses porcine epidemic diarrhea virus replication by targeting and degrading virus nucleocapsid protein with selective autophagy

BST2 suppresses porcine epidemic diarrhea virus replication by targeting and degrading virus nucleocapsid protein with selective autophagy

Identification of a novel B-cell epitope in the spike protein of porcine epidemic diarrhea virus

Tumor suppressor p53 inhibits porcine epidemic diarrhea virus infection via interferon-mediated antiviral immunity

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