The Network of Interactions between the Porcine Epidemic Diarrhea Virus Nucleocapsid and Host Cellular Proteins

Zhou, Jianwei; Qiu, Yonghui; Zhao, Jie; Wang, Yongxia; Zhu, Ning; Wang, Dedong; Cui, Yongqiu; Guo, Jinshuo; Sun, T.; Jin, Ying; Wu, Zhi Xue; Zeng, Peng-Hui; Li, Jingyi; Feng, Xufei; Hou, Lei; Liu, Jue

doi:10.3390/v14102269

Cited by 3 publications

(3 citation statements)

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“…These include mitochondrial translation, mRNA metabolic process, ribosome biogenesis, regulation of translation, ribonucleoprotein complex assembly, RNA localization, spliceosomal complex assembly, RNA modification, RNA 3’-end processing, among others (Fig 6A-B). Notably, these findings align with the previous interactome analyses of the N proteins from IBV [50], PEDV [51], SARS-CoV-2 [52, 53]. Among the IBV N binding proteins involved in nucleocytoplasmic transport, key players were identified and listed in Fig 6C, including RACK1 (Receptor of activated protein C kinase 1), NXF1 (Nuclear RNA export factor 1), LMNB1 (Lamin B1), KPNA2 (importin α1), PPP1CC (Protein phosphatase PP1γ), RAE1 (mRNA export factor), LBR (Lamin B receptor), SRPK1 (SRSF protein kinase 1), NUPL2 (Nucleoporin-like protein 2, NUP42).…”

Section: Resultssupporting

confidence: 89%

“…These include mitochondrial translation, mRNA metabolic process, ribosome biogenesis, regulation of translation, ribonucleoprotein complex assembly, RNA localization, spliceosomal complex assembly, RNA modification, RNA 3'-end processing, among others (Fig 6A -B). Notably, these findings align with the previous interactome analyses of the N proteins from IBV [50], PEDV [51], SARS-CoV-2 [52,53]. Among the IBV N binding proteins involved in nucleocytoplasmic transport, key To investigate whether the interaction with RACK1 is a common feature among pan-coronaviruses N proteins, Flag-tagged N proteins from various coronaviruses were co-expressed with HA-RACK1 in HEK-293T cells.…”

Section: Pan-coronaviruses N Proteins Interact With Scaffold Protein ...supporting

confidence: 80%

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Coronavirus Nucleocapsid Protein Enhances the binding of p-PKCα to RACK1: Implications for Inhibition of Nucleocytoplasmic Trafficking and Suppression of the Innate Immune Response

Xue,

Chu,

Wang

et al. 2024

Preprint

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The hallmark of coronavirus infection lies in its ability to evade host immune defenses, a process intricately linked to the nuclear entry of transcription factors crucial for initiating the expression of antiviral genes. Central to this evasion strategy is the manipulation of the nucleocytoplasmic trafficking system, which serves as an effective target for the virus to modulate the expression of immune response-related genes. In this investigation, we discovered that infection with the infectious bronchitis virus (IBV) dynamically impedes the nuclear translocation of several transcription factors such as IRF3, STAT1, STAT2, NF-κB p65, and the p38 mitogen-activated protein kinase (MAPK), leading to compromised transcriptional induction of key antiviral genes such as IFNβ, IFITM3, and IL-8. Further examination revealed that during the infection process, components of the nuclear pore complex (NPC), particularly FG-Nups (such as NUP62, NUP153, NUP42, and TPR), undergo cytosolic dispersion from the nuclear envelope; NUP62 undergoes phosphorylation, and NUP42 exhibits a mobility shift in size. These observations suggest a disruption in nucleocytoplasmic trafficking. Screening efforts identified the IBV nucleocapsid protein (N) as the agent responsible for the cytoplasmic distribution of FG-Nups, subsequently hindering the nuclear entry of transcription factors and suppressing the expression of antiviral genes. Interactome analysis further revealed that the IBV N protein interacts with the scaffold protein RACK1, facilitating the recruitment of activated protein kinase C alpha (p-PKCα) to RACK1 and relocating the RACK1-PKCα complex to the cytoplasm. These observations are conserved across pan-coronaviruses N proteins. Concurrently, the presence of both RACK1 and PKCα/β proved essential for the phosphorylation and cytoplasmic dispersion of NUP62, the suppression of antiviral cytokine expression, and efficient virus replication. These findings unveil a novel, highly effective, and evolutionarily conserved mechanism.Author summaryCoronaviruses employ diverse strategies to suppress the host innate immune defense. In this study, we uncovered a novel and highly effective strategy utilized by pan-coronaviruses to inhibit the innate immune response. Specifically, we found that the coronavirus N protein facilitates the binding of p-PKCα to RACK1, leading to the phosphorylation of NUP62 and the cytoplasmic redistribution of multiple FG-Nups. This phenomenon is accompanied by the disruption of nuclear translocation of several innate immune response-related transcription factors and suppression of antiviral/pro-inflammatory genes expression. Our research represents the first elucidation of how the N protein targets and impairs NPC function through the promotion of RACK1-PKCα interaction and NUP62 phosphorylation/disassembly. This discovery unveils a novel mechanism employed by pan-coronaviruses to counteract the host immune response.

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Section: Resultssupporting

confidence: 89%

“…These include mitochondrial translation, mRNA metabolic process, ribosome biogenesis, regulation of translation, ribonucleoprotein complex assembly, RNA localization, spliceosomal complex assembly, RNA modification, RNA 3'-end processing, among others (Fig 6A -B). Notably, these findings align with the previous interactome analyses of the N proteins from IBV [50], PEDV [51], SARS-CoV-2 [52,53]. Among the IBV N binding proteins involved in nucleocytoplasmic transport, key To investigate whether the interaction with RACK1 is a common feature among pan-coronaviruses N proteins, Flag-tagged N proteins from various coronaviruses were co-expressed with HA-RACK1 in HEK-293T cells.…”

Section: Pan-coronaviruses N Proteins Interact With Scaffold Protein ...supporting

confidence: 80%

Coronavirus Nucleocapsid Protein Enhances the binding of p-PKCα to RACK1: Implications for Inhibition of Nucleocytoplasmic Trafficking and Suppression of the Innate Immune Response

Xue,

Chu,

Wang

et al. 2024

Preprint

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“…The DDX21 -knockdown PK-15 cells were constructed as previously described ( Zhou et al, 2022a ). All media were supplemented with 10% fetal bovine serum (S711-001S; LONSERA, Shanghai Shuangru Biology Science & Technology Co., Ltd, China) as previously described ( Zhou et al, 2022c ). PCV2 strain BJW (accession no.…”

Section: Methodsmentioning

confidence: 99%

DEAD-box RNA helicase 21 interacts with porcine circovirus type 2 Cap protein and facilitates viral replication

Zhou,

Zhao,

Sun

et al. 2024

Front. Microbiol.

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Porcine circovirus type 2 (PCV2) is the etiological agent of PCV2-associated diseases that pose a serious threat to the swine industry. PCV2 capsid (Cap) protein has been shown to interact with DEAD-box RNA helicase 21 (DDX21), an important protein that regulates RNA virus replication. However, whether the interaction between DDX21 and the PCV2 Cap regulates PCV2 replication remains unclear. Herein, by using western blotting, interaction assays, and knockdown analysis, we found that PCV2 infection induced the cytoplasmic relocation of DDX21 from the nucleolus in cultured PK-15 cells. Moreover, the nuclear localization signal (NLS) of PCV2 Cap interacted directly with DDX21. The NLS of PCV2 Cap and 763GSRSNRFQNK772 residues at the C-terminal domain (CTD) of DDX21 were essential for the dual interaction. Upon shRNA-mediated DDX21 depletion in PK-15 cells, we observed impaired PCV2 replication via a lentivirus-delivered system, as evidenced by decreased levels of viral protein expression and virus production. In contrast, the replication of PCV2 increased in transiently DDX21-overexpressing cells. Our results indicate that DDX21 interacts with PCV2 Cap and plays a crucial role in virus replication. These results provide a reference for developing novel potential targets for prevention and control of PCV2 infection.

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Phylogenetic and Evolutionary Analysis of Porcine Epidemic Diarrhea Virus in Guangxi Province, China, during 2020 and 2024

Shi,

Li,

Shi

et al. 2024

Viruses

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The variant porcine epidemic diarrhea virus (PEDV) has caused considerable economic losses to the global pig industry since 2010. In this study, a total of 5859 diarrhea samples were collected from different pig farms in China’s Guangxi province during January 2020 and March 2024 and tested for PEDV using RT-qPCR. The positivity rate of PEDV was 11.90% (697/5859). Ninety-two PEDV-positive samples were selected based on sampling time, and the sampling region for amplification, sequencing, and analysis of the S1, M, and N genes. Phylogenetic analysis of the S1 gene revealed that all strains from Guangxi province were distributed in three subgroups, i.e., 81.5% (75/92) in the G2a subgroup, 4.3% (4/92) in the G2b subgroup, and 14.1% (13/92) in the G2c subgroup. The sequence analysis revealed that the S1 gene sequences from Guangxi province had higher homology with the variant strains than with the classical strains, showing as high as 99.2% with the variant strain AJ1102 and only 94.3% with the classical strain CV777. Recombination analysis revealed that the GX-BS08-2023 strain (G2c) from Guangxi province originated from inter-lineage recombination between the GX-BS09-2023 (G2a) and CH-JN547228-2011 (G1a) strains. In addition, the S1 gene of the G2a and G2b subgroup strains shared many mutations and insertions. There were common mutations of N143D and P235L in the G2a subgroup. Evolutionary analysis revealed that all Guangxi strains belonged to the G2 genotype. These strains have spread rapidly since the PEDV variant strains that emerged in 2010, weakened until 2021, and then remained stable. In conclusion, the results revealed the latest genetic evolution of circulating PEDV strains in Guangxi province in recent years, providing important information for preventing and controlling PEDV infection. Currently, the G2a subgroup strains are the predominant strains circulating in pig herds in Guangxi province, southern China.

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The Network of Interactions between the Porcine Epidemic Diarrhea Virus Nucleocapsid and Host Cellular Proteins

Cited by 3 publications

References 50 publications

Coronavirus Nucleocapsid Protein Enhances the binding of p-PKCα to RACK1: Implications for Inhibition of Nucleocytoplasmic Trafficking and Suppression of the Innate Immune Response

Coronavirus Nucleocapsid Protein Enhances the binding of p-PKCα to RACK1: Implications for Inhibition of Nucleocytoplasmic Trafficking and Suppression of the Innate Immune Response

DEAD-box RNA helicase 21 interacts with porcine circovirus type 2 Cap protein and facilitates viral replication

Phylogenetic and Evolutionary Analysis of Porcine Epidemic Diarrhea Virus in Guangxi Province, China, during 2020 and 2024

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