Porcine Deltacoronavirus (PDCoV) Entry into PK-15 Cells by Caveolae-Mediated Endocytosis

• Published in: Viruses Vol. 14; no. 3; p. 496
• Main Authors: Li, Shiqian, Xiao, Dai, Zhao, Yujia, Zhang, Luwen, Chen, Rui, Liu, Weizhe, Wen, Yimin, Liao, Yijie, Wen, Yiping, Wu, Rui, Han, Xinfeng, Zhao, Qin, Du, Senyan, Yan, Qigui, Wen, Xintian, Cao, Sanjie, Huang, Xiaobo
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 28.02.2022; MDPI
• Subjects: Acidification; Amiloride; Ammonium; Ammonium chloride; Animals; Antibodies; Caveolae; Caveolae - metabolism; caveolae-mediated endocytosis; cell entry; Cell Line; Chlorpromazine; Cholesterol; Cholesterol - metabolism; Clathrin; Clathrin - metabolism; clathrin-mediated endocytosis; Coronaviruses; Deltacoronavirus; Dynamin; Dynamins - metabolism; Endocytosis; Epidemics; Immunofluorescence; Internalization; macropinocytosis; Membranes; Methyl-β-Cyclodextrin; Nystatin; Plasmids; porcine deltacoronavirus (PDCoV); Proteins; Severe acute respiratory syndrome coronavirus 2; Swine; Virus Internalization; Viruses; United States > US; China; Germany; cell entry; caveolae-mediated endocytosis; macropinocytosis; porcine deltacoronavirus (PDCoV); clathrin-mediated endocytosis
• ISSN: 1999-4915; 1999-4915
• DOI: 10.3390/v14030496

(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 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.