Unveiling the Role of Protein Kinase C θ in Porcine Epidemic Diarrhea Virus Replication: Insights from Genome-Wide CRISPR/Cas9 Library Screening

被引：4

作者：

Zhou, Jinglin ^{[1
]}

Feng, Zhihua ^{[1
]}

Lv, Deyang ^{[1
]}

Wang, Duokai ^{[1
]}

Sang, Kai ^{[1
]}

Liu, Zhihao ^{[1
]}

Guo, Dong ^{[1
]}

Shen, Yangkun ^{[1
]}

Chen, Qi ^{[1
]}

机构：

[1] Fujian Normal Univ, Coll Life Sci, Biomed Res Ctr South China, Fujian Key Lab Innate Immune Biol, Qishan Campus, Fuzhou 350117, Peoples R China

来源：

INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES | 2024年 / 25卷 / 06期

关键词：

genome-scale CRISPR screen; PEDV (porcine epidemic diarrhea virus); susceptibility; PKC theta (protein kinase C theta); BOK (B-cell lymphoma 2 (BCL-2) ovarian killer); mitochondrial apoptosis; PEDV endocytosis; PEDV replication; INTESTINAL EPITHELIAL-CELLS; RESPIRATORY SYNDROME VIRUS; PKC-THETA; INFECTION; ACTIVATION; OUTBREAK; DEATH; SWINE; APOPTOSIS;

D O I：

10.3390/ijms25063096

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Porcine epidemic diarrhea virus (PEDV), a member of the Alpha-coronavirus genus in the Coronaviridae family, induces acute diarrhea, vomiting, and dehydration in neonatal piglets. This study aimed to investigate the genetic dependencies of PEDV and identify potential therapeutic targets by using a single-guide RNA (sgRNA) lentiviral library to screen host factors required for PEDV infection. Protein kinase C theta (PKC theta), a calcium-independent member of the PKC family localized in the cell membrane, was found to be a crucial host factor in PEDV infection. The investigation of PEDV infection was limited in Vero and porcine epithelial cell-jejunum 2 (IPEC-J2) due to defective interferon production in Vero and the poor replication of PEDV in IPEC-J2. Therefore, identifying suitable cells for PEDV investigation is crucial. The findings of this study reveal that human embryonic kidney (HEK) 293T and L929 cells, but not Vero and IPEC-J2 cells, were suitable for investigating PEDV infection. PKC theta played a significant role in endocytosis and the replication of PEDV, and PEDV regulated the expression and phosphorylation of PKC theta. Apoptosis was found to be involved in PEDV replication, as the virus activated the PKC theta-B-cell lymphoma 2 (BCL-2) ovarian killer (BOK) axis in HEK293T and L929 cells to increase viral endocytosis and replication via mitochondrial apoptosis. This study demonstrated the suitability of HEK293T and L929 cells for investigating PEDV infection and identified PKC theta as a host factor essential for PEDV infection. These findings provide valuable insights for the development of strategies and drug targets for PEDV infection.

引用

页数：27

共 53 条

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