Unveiling the Role of Protein Kinase C θ in Porcine Epidemic Diarrhea Virus Replication: Insights from Genome-Wide CRISPR/Cas9 Library Screening
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 s...
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MDPI AG
2024-03-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/25/6/3096 |
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| author | Jinglin Zhou Zhihua Feng Deyang Lv Duokai Wang Kai Sang Zhihao Liu Dong Guo Yangkun Shen Qi Chen |
| author_facet | Jinglin Zhou Zhihua Feng Deyang Lv Duokai Wang Kai Sang Zhihao Liu Dong Guo Yangkun Shen Qi Chen |
| author_sort | Jinglin Zhou |
| collection | DOAJ |
| description | 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 θ (PKCθ), 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. PKCθ played a significant role in endocytosis and the replication of PEDV, and PEDV regulated the expression and phosphorylation of PKCθ. Apoptosis was found to be involved in PEDV replication, as the virus activated the PKCθ-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 PKCθ as a host factor essential for PEDV infection. These findings provide valuable insights for the development of strategies and drug targets for PEDV infection. |
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| language | English |
| last_indexed | 2024-04-24T18:12:04Z |
| publishDate | 2024-03-01 |
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| spelling | doaj.art-200f10416f5749ae866bfaa815d737e12024-03-27T13:45:00ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672024-03-01256309610.3390/ijms25063096Unveiling the Role of Protein Kinase C θ in Porcine Epidemic Diarrhea Virus Replication: Insights from Genome-Wide CRISPR/Cas9 Library ScreeningJinglin Zhou0Zhihua Feng1Deyang Lv2Duokai Wang3Kai Sang4Zhihao Liu5Dong Guo6Yangkun Shen7Qi Chen8Fujian Key Laboratory of Innate Immune Biology, Biomedical Research Center of South China, College of Life Science, Qishan Campus, Fujian Normal University, Fuzhou 350117, ChinaFujian Key Laboratory of Innate Immune Biology, Biomedical Research Center of South China, College of Life Science, Qishan Campus, Fujian Normal University, Fuzhou 350117, ChinaFujian Key Laboratory of Innate Immune Biology, Biomedical Research Center of South China, College of Life Science, Qishan Campus, Fujian Normal University, Fuzhou 350117, ChinaFujian Key Laboratory of Innate Immune Biology, Biomedical Research Center of South China, College of Life Science, Qishan Campus, Fujian Normal University, Fuzhou 350117, ChinaFujian Key Laboratory of Innate Immune Biology, Biomedical Research Center of South China, College of Life Science, Qishan Campus, Fujian Normal University, Fuzhou 350117, ChinaFujian Key Laboratory of Innate Immune Biology, Biomedical Research Center of South China, College of Life Science, Qishan Campus, Fujian Normal University, Fuzhou 350117, ChinaFujian Key Laboratory of Innate Immune Biology, Biomedical Research Center of South China, College of Life Science, Qishan Campus, Fujian Normal University, Fuzhou 350117, ChinaFujian Key Laboratory of Innate Immune Biology, Biomedical Research Center of South China, College of Life Science, Qishan Campus, Fujian Normal University, Fuzhou 350117, ChinaFujian Key Laboratory of Innate Immune Biology, Biomedical Research Center of South China, College of Life Science, Qishan Campus, Fujian Normal University, Fuzhou 350117, ChinaPorcine 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 θ (PKCθ), 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. PKCθ played a significant role in endocytosis and the replication of PEDV, and PEDV regulated the expression and phosphorylation of PKCθ. Apoptosis was found to be involved in PEDV replication, as the virus activated the PKCθ-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 PKCθ as a host factor essential for PEDV infection. These findings provide valuable insights for the development of strategies and drug targets for PEDV infection.https://www.mdpi.com/1422-0067/25/6/3096genome-scale CRISPR screenPEDV (porcine epidemic diarrhea virus)susceptibilityPKCθ (protein kinase C θ)BOK (B-cell lymphoma 2 (BCL-2) ovarian killer)mitochondrial apoptosis |
| spellingShingle | Jinglin Zhou Zhihua Feng Deyang Lv Duokai Wang Kai Sang Zhihao Liu Dong Guo Yangkun Shen Qi Chen Unveiling the Role of Protein Kinase C θ in Porcine Epidemic Diarrhea Virus Replication: Insights from Genome-Wide CRISPR/Cas9 Library Screening International Journal of Molecular Sciences genome-scale CRISPR screen PEDV (porcine epidemic diarrhea virus) susceptibility PKCθ (protein kinase C θ) BOK (B-cell lymphoma 2 (BCL-2) ovarian killer) mitochondrial apoptosis |
| title | Unveiling the Role of Protein Kinase C θ in Porcine Epidemic Diarrhea Virus Replication: Insights from Genome-Wide CRISPR/Cas9 Library Screening |
| title_full | Unveiling the Role of Protein Kinase C θ in Porcine Epidemic Diarrhea Virus Replication: Insights from Genome-Wide CRISPR/Cas9 Library Screening |
| title_fullStr | Unveiling the Role of Protein Kinase C θ in Porcine Epidemic Diarrhea Virus Replication: Insights from Genome-Wide CRISPR/Cas9 Library Screening |
| title_full_unstemmed | Unveiling the Role of Protein Kinase C θ in Porcine Epidemic Diarrhea Virus Replication: Insights from Genome-Wide CRISPR/Cas9 Library Screening |
| title_short | Unveiling the Role of Protein Kinase C θ in Porcine Epidemic Diarrhea Virus Replication: Insights from Genome-Wide CRISPR/Cas9 Library Screening |
| title_sort | unveiling the role of protein kinase c θ in porcine epidemic diarrhea virus replication insights from genome wide crispr cas9 library screening |
| topic | genome-scale CRISPR screen PEDV (porcine epidemic diarrhea virus) susceptibility PKCθ (protein kinase C θ) BOK (B-cell lymphoma 2 (BCL-2) ovarian killer) mitochondrial apoptosis |
| url | https://www.mdpi.com/1422-0067/25/6/3096 |
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