Dependence of SARS-CoV-2 infection on cholesterol-rich lipid raft and endosomal acidification

  • Comput Struct Biotechnol J. 2021;19:1933-1943. doi: 10.1016/j.csbj.2021.04.001.
Xiaowei Li  1  2  3 ,  Wenhua Zhu  2  3 ,  Meiyang Fan  2  3 ,  Jing Zhang  2  3 ,  Yizhao Peng  2  3 ,  Fumeng Huang  2  3 ,  Nan Wang  4 ,  Langchong He  4 ,  Lei Zhang  5 ,  Rikard Holmdahl  1  2  3  6 ,  Liesu Meng  1  2  3 ,  Shemin Lu  1  2  3
Affiliations
  • 1. National Joint Engineering Research Center of Biodiagnostics and Biotherapy, Second Affiliated Hospital, Xi'an Jiaotong University, 710004 Xi'an, China.
  • 2. Institute of Molecular and Translational Medicine (IMTM), and Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Xi'an Jiaotong University Health Science Center, 710061 Xi'an, China.
  • 3. Key Laboratory of Environment and Genes Related to Diseases (Xi'an Jiaotong University), Ministry of Education of China, 710061 Xi'an, China.
  • 4. School of Pharmacy, Xi'an Jiaotong University, 710061 Xi'an, China.
  • 5. Shaanxi Provincial Centre for Disease Control and Prevention, 710054 Xi'an, China.
  • 6. Section for Medical Inflammation Research, Department of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm 171 77, Sweden.
PMID: 33850607 DOI: 10.1016/j.csbj.2021.04.001
Abstract

Coronavirus disease 2019 is a kind of viral pneumonia caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). However, the mechanism whereby SARS-CoV-2 invades host cells remains poorly understood. Here we used SARS-CoV-2 pseudoviruses to infect human angiotensin-converting enzyme 2 (ACE2) expressing HEK293T cells and evaluated virus Infection. We confirmed that SARS-CoV-2 entry was dependent on ACE2 and sensitive to pH of endosome/lysosome in HEK293T cells. The Infection of SARS-CoV-2 pseudoviruses is independent of Dynamin, clathrin, caveolin and endophilin A2, as well as macropinocytosis. Instead, we found that the Infection of SARS-CoV-2 pseudoviruses was cholesterol-rich lipid raft dependent. Cholesterol depletion of cell membranes with methyl-β-cyclodextrin resulted in reduction of pseudovirus Infection. The Infection of SARS-CoV-2 pseudoviruses resumed with Cholesterol supplementation. Together, cholesterol-rich lipid rafts, and endosomal acidification, are key steps of SARS-CoV-2 required for Infection of host cells. Therefore, our finding expands the understanding of SARS-CoV-2 entry mechanism and provides a new anti-SARS-CoV-2 strategy.

Keywords
Cholesterol; Endocytosis; Endosomal acidification; Lipid rafts; SARS-CoV-2.
Products