SARS-CoV-2 pseudovirus enters the host cells through spike protein-CD147 in an Arf6-dependent manner

Abstract

The spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its variants is threatening public health around the world. Endocytosis functions as an important way for viral infection, and SARS-CoV-2 bears no exception. However, the specific endocytic mechanism of SARS-CoV-2 remains unknown. In this study, we used endocytic inhibitors to evaluate the role of different endocytic routes in SARS-CoV-2 pseudovirus infection and found that the viral infection was associated with caveolar/lipid raft- and cytoskeleton-mediated endocytosis, but independent of the clathrin-mediated endocytosis and macropinocytosis. Meanwhile, the knockdown of CD147 and Rab5a in Vero E6 and Huh-7 cells inhibited SARS-CoV-2 pseudovirus infection, and the co-localization of spike protein, CD147, and Rab5a was observed in pseudovirus-infected Vero E6 cells, which was weakened by CD147 silencing, illustrating that SARS-CoV-2 pseudovirus entered the host cells via CD147-mediated endocytosis. Additionally, Arf6 silencing markedly inhibited pseudovirus infection in Vero E6 and Huh-7 cells, while little change was observed in CD147 knockout-Vero E6 cells. This finding indicated Arf6-mediated CD147 trafficking plays a vital role in SARS-CoV-2 entry. Taken together, our findings provide new insights into the CD147-Arf6 axis in mediating SARS-CoV-2 pseudovirus entry into the host cells, and further suggest that blockade of this pathway seems to be a feasible approach to prevent the SARS-CoV-2 infection clinically.

ARF6 is a host factor for SARS-CoV-2 infectionin vitro

“…Thus, differences in virus and/or cell type may alter the endocytic uptake pathway. During the revision of this manuscript, Zhou et al also published that ARF6 is an important host factor for SARS-CoV-2 entry [20]. Specifically, their study used an siRNA approach and SARS-CoV-2 pseudovirus to demonstrate a 2-fold reduction of viral entry in Huh-7 cells.…”

Endomembrane remodeling in SARS-CoV-2 infection

“…In the absence of TMPRSS2, the ACE2-engaged SARS-CoV-2 virion is internalized into the cytosol through clathrin-mediated endocytosis ( Bayati et al, 2021 ; Yang et al, 2022 ). SARS-CoV-2 pseudovirus infection assays suggest that caveolar/lipid raft- and cytoskeleton-mediated endocytosis are also involved in viral infection ( Li et al, 2021a ; Zhou et al, 2022 ). After fusion of virion-containing endosomes with lysosomes, S2 subunit is cleaved by cathepsin cysteine proteases (e.g.…”

Cyclophilin A/CD147 Interaction: A Promising Target for Anticancer Therapy

“…The inhibition of CD147 expression alleviated atherosclerosis and exhibited therapeutic effects in collagen-induced arthritis [ 45 , 46 ]. CD147 reportedly serves as a receptor for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) [ 47 ]. Furthermore, CD147 is overexpressed in various cancer cells and promotes the proliferation, metastasis, angiogenesis, and stemness of cancer cells through the activation of major oncogenic signaling pathways [ 15 , 44 , 48 , 49 ].…”