Sequestration of membrane cholesterol by cholesterol-binding proteins inhibits SARS-CoV-2 entry into Vero E6 cells

Kulma, Magdalena (author), Šakanović, Aleksandra (author), Bedina Zavec, Apolonija (author), Caserman, Simon (author), Omersa, Neža (author), Šolinc, Gašper (author), Orehek, Sara (author), Hafner Bratkovič, Iva (author), Kuhar, Urška (author), Slavec, Brigita (author), Krapež, Uroš (author), Ocepek, Matjaž (author), Kobayashi, Toshihide (author), Kwiatkowska, Katarzyna (author), Jerala, Roman (author), Podobnik, Marjetka (author), Anderluh, Gregor (author)

Membrane lipids and proteins form dynamic domains crucial for physiological and pathophysiological processes, including viral infection. Many plasma membrane proteins, residing within membrane domains enriched with cholesterol (CHOL) and sphingomyelin (SM), serve as receptors for attachment and entry of viruses into the host cell. Among these, human coronaviruses, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), use proteins associated with membrane domains for initial binding and internalization. We hypothesized that the interaction of lipid-binding proteins with CHOL in plasma membrane could sequestrate lipids and thus affect the efficiency of virus entry into host cells, preventing the initial steps of viral infection. We have prepared CHOL-binding proteins with high affinities for lipids in the plasma membrane of mammalian cells. Binding of the perfringolysin O domain four (D4) and its variant D4$^{E458L}$ to membrane CHOL impaired the internalization of the receptor-binding domain of the SARS-CoV-2 spike protein and the pseudovirus complemented with the SARS-CoV-2 spike protein. SARS-CoV-2 replication in Vero E6 cells was also decreased. Overall, our results demonstrate that the integrity of CHOL-rich membrane domains and the accessibility of CHOL in the membrane play an essential role in SARS-CoV-2 cell entry.

• Language: English
• Keywords: cholesterol, endocytosis, plasma membrane, membrane domains, pore-forming proteins, SARS-CoV-2, viral infection
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: VF - Veterinary Faculty
• Publication status: Published
• Publication version: Version of Record
• Year: 2024
• Number of pages: 16 str.
• Numbering: Vol. 716, art. 149954
• PID: 20.500.12556/RUL-156325
• UDC: 577
• ISSN on article: 1090-2104
• DOI: 10.1016/j.bbrc.2024.149954
• COBISS.SI-ID: 194174467
• Publication date in RUL: 20.05.2024

Record is a part of a journal

• Title: Biochemical and biophysical research communications
• Shortened title: Biochem. biophys. res. commun.
• Publisher: Elsevier
• ISSN: 1090-2104
• COBISS.SI-ID: 18258983

Licences

• License: CC BY-NC-ND 4.0, Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
• Link: http://creativecommons.org/licenses/by-nc-nd/4.0/
• Description: The most restrictive Creative Commons license. This only allows people to download and share the work for no commercial gain and for no other purposes.

Secondary language

• Language: Slovenian
• Keywords: biokemija, SARS-CoV-2, holesterol, endocitoza, beljakovine, koronavirusi

Projects

• Funder: ARIS - Slovenian Research and Innovation Agency
• Project number: P1-0391
• Name: Molekulske interakcije

• Funder: ARIS - Slovenian Research and Innovation Agency
• Project number: P4-0092
• Name: Zdravje živali, okolje in varna hrana