Subject(s)
COVID-19 , DNA Helicases , RNA Helicases , RNA Recognition Motif Proteins , SARS-CoV-2 , Virus Replication , Humans , Virus Replication/physiology , SARS-CoV-2/physiology , COVID-19/metabolism , COVID-19/virology , RNA Recognition Motif Proteins/metabolism , RNA Recognition Motif Proteins/physiology , RNA Helicases/metabolism , RNA Helicases/physiology , DNA Helicases/metabolism , DNA Helicases/physiology , Animals , Poly-ADP-Ribose Binding ProteinsABSTRACT
Considerable progress has been made in understanding the molecular host-virus battlefield during SARS-CoV-2 infection. Nevertheless, the assembly and egress of newly formed virions are less understood. To identify host proteins involved in viral morphogenesis, we characterize the proteome of SARS-CoV-2 virions produced from A549-ACE2 and Calu-3 cells, isolated via ultracentrifugation on sucrose cushion or by ACE-2 affinity capture. Bioinformatic analysis unveils 92 SARS-CoV-2 virion-associated host factors, providing a valuable resource to better understand the molecular environment of virion production. We reveal that G3BP1 and G3BP2 (G3BP1/2), two major stress granule nucleators, are embedded within virions and unexpectedly favor virion production. Furthermore, we show that G3BP1/2 participate in the formation of cytoplasmic membrane vesicles, that are likely virion assembly sites, consistent with a proviral role of G3BP1/2 in SARS-CoV-2 dissemination. Altogether, these findings provide new insights into host factors required for SARS-CoV-2 assembly with potential implications for future therapeutic targeting.