Porcine Deltacoronavirus (PDCoV) Entry into PK-15 Cells by Caveolae-Mediated Endocytosis
(1) Background: Porcine deltacoronavirus (PDCoV) is a newly emerged enteric virus affecting pig breeding industries worldwide, and its pathogenic mechanism remains unclear. (2) Methods: In this study, we preliminarily identified the endocytic pathway of PDCoV in PK-15 cells, using six chemical inhib...
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| Language: | English |
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MDPI AG
2022-02-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/14/3/496 |
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| author | Shiqian Li Dai Xiao Yujia Zhao Luwen Zhang Rui Chen Weizhe Liu Yimin Wen Yijie Liao Yiping Wen Rui Wu Xinfeng Han Qin Zhao Senyan Du Qigui Yan Xintian Wen Sanjie Cao Xiaobo Huang |
| author_facet | Shiqian Li Dai Xiao Yujia Zhao Luwen Zhang Rui Chen Weizhe Liu Yimin Wen Yijie Liao Yiping Wen Rui Wu Xinfeng Han Qin Zhao Senyan Du Qigui Yan Xintian Wen Sanjie Cao Xiaobo Huang |
| author_sort | Shiqian Li |
| collection | DOAJ |
| description | (1) Background: Porcine deltacoronavirus (PDCoV) is a newly emerged enteric virus affecting pig breeding industries worldwide, and its pathogenic mechanism remains unclear. (2) Methods: In this study, we preliminarily identified the endocytic pathway of PDCoV in PK-15 cells, using six chemical inhibitors (targeting clathrin-mediated endocytosis, caveolae-mediated endocytosis, macropinocytosis pathway and endosomal acidification), overexpression of dominant-negative (DN) mutants to treat PK-15 cells and proteins knockdown. (3) Results: The results revealed that PDCoV entry was not affected after treatment with chlorpromazine (CPZ), 5-(N-ethyl-N-isopropyl) amiloride (EIPA)or ammonium chloride (NH<sub>4</sub>Cl), indicating that the entry of PDCoV into PK-15 cells were clathrin-, micropinocytosis-, PH-independent endocytosis. Conversely, PDCoV infection was sensitive to nystatin, dynasore and methyl-β-cyclodextrin (MβCD) with reduced PDCoV internalization, indicating that entry of PDCoV into PK-15 cells was caveolae-mediated endocytosis that required dynamin and cholesterol; indirect immunofluorescence and shRNA interference further validated these results. (4) Conclusions: In conclusion, PDCoV entry into PK-15 cells depends on caveolae-mediated endocytosis, which requires cholesterol and dynamin. Our finding is the first initial identification of the endocytic pathway of PDCoV in PK-15 cells, providing a theoretical basis for an in-depth understanding of the pathogenic mechanism of PDCoV and the design of new antiviral targets. |
| first_indexed | 2024-03-09T12:17:17Z |
| format | Article |
| id | doaj.art-e873a3eea3d94f4486a8c8363b223c1d |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-09T12:17:17Z |
| publishDate | 2022-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-e873a3eea3d94f4486a8c8363b223c1d2023-11-30T22:45:24ZengMDPI AGViruses1999-49152022-02-0114349610.3390/v14030496Porcine Deltacoronavirus (PDCoV) Entry into PK-15 Cells by Caveolae-Mediated EndocytosisShiqian Li0Dai Xiao1Yujia Zhao2Luwen Zhang3Rui Chen4Weizhe Liu5Yimin Wen6Yijie Liao7Yiping Wen8Rui Wu9Xinfeng Han10Qin Zhao11Senyan Du12Qigui Yan13Xintian Wen14Sanjie Cao15Xiaobo Huang16Research Center for Swine Diseases, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, ChinaResearch Center for Swine Diseases, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, ChinaResearch Center for Swine Diseases, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, ChinaResearch Center for Swine Diseases, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, ChinaResearch Center for Swine Diseases, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, ChinaResearch Center for Swine Diseases, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, ChinaResearch Center for Swine Diseases, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, ChinaResearch Center for Swine Diseases, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, ChinaResearch Center for Swine Diseases, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, ChinaResearch Center for Swine Diseases, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, ChinaResearch Center for Swine Diseases, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, ChinaResearch Center for Swine Diseases, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, ChinaResearch Center for Swine Diseases, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, ChinaResearch Center for Swine Diseases, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, ChinaResearch Center for Swine Diseases, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, ChinaResearch Center for Swine Diseases, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, ChinaResearch Center for Swine Diseases, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China(1) Background: Porcine deltacoronavirus (PDCoV) is a newly emerged enteric virus affecting pig breeding industries worldwide, and its pathogenic mechanism remains unclear. (2) Methods: In this study, we preliminarily identified the endocytic pathway of PDCoV in PK-15 cells, using six chemical inhibitors (targeting clathrin-mediated endocytosis, caveolae-mediated endocytosis, macropinocytosis pathway and endosomal acidification), overexpression of dominant-negative (DN) mutants to treat PK-15 cells and proteins knockdown. (3) Results: The results revealed that PDCoV entry was not affected after treatment with chlorpromazine (CPZ), 5-(N-ethyl-N-isopropyl) amiloride (EIPA)or ammonium chloride (NH<sub>4</sub>Cl), indicating that the entry of PDCoV into PK-15 cells were clathrin-, micropinocytosis-, PH-independent endocytosis. Conversely, PDCoV infection was sensitive to nystatin, dynasore and methyl-β-cyclodextrin (MβCD) with reduced PDCoV internalization, indicating that entry of PDCoV into PK-15 cells was caveolae-mediated endocytosis that required dynamin and cholesterol; indirect immunofluorescence and shRNA interference further validated these results. (4) Conclusions: In conclusion, PDCoV entry into PK-15 cells depends on caveolae-mediated endocytosis, which requires cholesterol and dynamin. Our finding is the first initial identification of the endocytic pathway of PDCoV in PK-15 cells, providing a theoretical basis for an in-depth understanding of the pathogenic mechanism of PDCoV and the design of new antiviral targets.https://www.mdpi.com/1999-4915/14/3/496porcine deltacoronavirus (PDCoV)clathrin-mediated endocytosiscaveolae-mediated endocytosismacropinocytosiscell entry |
| spellingShingle | Shiqian Li Dai Xiao Yujia Zhao Luwen Zhang Rui Chen Weizhe Liu Yimin Wen Yijie Liao Yiping Wen Rui Wu Xinfeng Han Qin Zhao Senyan Du Qigui Yan Xintian Wen Sanjie Cao Xiaobo Huang Porcine Deltacoronavirus (PDCoV) Entry into PK-15 Cells by Caveolae-Mediated Endocytosis Viruses porcine deltacoronavirus (PDCoV) clathrin-mediated endocytosis caveolae-mediated endocytosis macropinocytosis cell entry |
| title | Porcine Deltacoronavirus (PDCoV) Entry into PK-15 Cells by Caveolae-Mediated Endocytosis |
| title_full | Porcine Deltacoronavirus (PDCoV) Entry into PK-15 Cells by Caveolae-Mediated Endocytosis |
| title_fullStr | Porcine Deltacoronavirus (PDCoV) Entry into PK-15 Cells by Caveolae-Mediated Endocytosis |
| title_full_unstemmed | Porcine Deltacoronavirus (PDCoV) Entry into PK-15 Cells by Caveolae-Mediated Endocytosis |
| title_short | Porcine Deltacoronavirus (PDCoV) Entry into PK-15 Cells by Caveolae-Mediated Endocytosis |
| title_sort | porcine deltacoronavirus pdcov entry into pk 15 cells by caveolae mediated endocytosis |
| topic | porcine deltacoronavirus (PDCoV) clathrin-mediated endocytosis caveolae-mediated endocytosis macropinocytosis cell entry |
| url | https://www.mdpi.com/1999-4915/14/3/496 |
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