Glycosylation and S-palmitoylation regulate SARS-CoV-2 spike protein intracellular trafficking
Chih-Feng Tien,
Wan-Ting Tsai,
Chun Hwa Chen,
Hui-Ju Chou,
Mingzi M. Zhang,
Jhe-Jhih Lin,
En-Ju Lin,
Shih-Syong Dai,
Yueh-Hsin Ping,
Chia-Yi Yu,
Yi-Ping Kuo,
Wei-Hsiang Tsai,
Hsin-Wei Chen,
Guann-Yi Yu
Affiliations
Chih-Feng Tien
National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, Zhunan, Taiwan
Wan-Ting Tsai
National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, Zhunan, Taiwan
Chun Hwa Chen
Institute of Molecular and Genomic Medicine, National Health Research Institutes, Zhunan, Taiwan
Hui-Ju Chou
Institute of Molecular and Genomic Medicine, National Health Research Institutes, Zhunan, Taiwan
Mingzi M. Zhang
Institute of Molecular and Genomic Medicine, National Health Research Institutes, Zhunan, Taiwan
Jhe-Jhih Lin
National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, Zhunan, Taiwan
En-Ju Lin
National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, Zhunan, Taiwan
Shih-Syong Dai
National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, Zhunan, Taiwan
Yueh-Hsin Ping
Department and Institute of Pharmacology, School of Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan
Chia-Yi Yu
National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, Zhunan, Taiwan
Yi-Ping Kuo
National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, Zhunan, Taiwan
Wei-Hsiang Tsai
National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, Zhunan, Taiwan
Hsin-Wei Chen
National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, Zhunan, Taiwan; Graduate Institute of Biomedical Sciences, China Medical University, Taichung, Taiwan; Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan
Guann-Yi Yu
National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, Zhunan, Taiwan; Corresponding author
Summary: Post-translational modifications (PTMs), such as glycosylation and palmitoylation, are critical to protein folding, stability, intracellular trafficking, and function. Understanding regulation of PTMs of SARS-CoV-2 spike (S) protein could help the therapeutic drug design. Herein, the VSV vector was used to produce SARS-CoV-2 S pseudoviruses to examine the roles of the 611LYQD614 and cysteine-rich motifs in S protein maturation and virus infectivity. Our results show that 611LY612 mutation alters S protein intracellular trafficking and reduces cell surface expression level. It also changes S protein glycosylation pattern and decreases pseudovirus infectivity. The S protein contains four cysteine-rich clusters with clusters I and II as the main palmitoylation sites. Mutations of clusters I and II disrupt S protein trafficking from ER-to-Golgi, suppress pseudovirus production, and reduce spike-mediated membrane fusion activity. Taken together, glycosylation and palmitoylation orchestrate the S protein maturation processing and are critical for S protein-mediated membrane fusion and infection.
