The global spread of SARS-CoV-2 is posing major public health challenges. One feature of SARS-CoV-2 spike protein is the insertion of multi-basic residues at the S1/S2 subunit cleavage site. Here, we find that the virus with intact spike (Sfull) preferentially enters cells via fusion at the plasma membrane, whereas a clone (Sdel) with deletion disrupting the multi-basic S1/S2 site utilizes an endosomal entry pathway. Using Sdel as model, we perform a genome-wide CRISPR screen and identify several endosomal entry-specific regulators. Experimental validation of hits from the CRISPR screen shows that host factors regulating the surface expression of angiotensin-converting enzyme 2 (ACE2) affect entry of Sfull virus. Animal-to-animal transmission with the Sdel virus is reduced compared to Sfull in the hamster model. These findings highlight the critical role of the S1/S2 boundary of SARS-CoV-2 spike protein in modulating virus entry and transmission and provide insights into entry of coronaviruses. The SARS-CoV-2 spike protein contains a multi-basic cleavage site. Here, the authors show how this multi-basic cleavage site affects entry of SARS-CoV-2 into cells and transmission in the hamster model and identify host factors affecting entry of SARS-CoV-2 in a genome-wide CRISPR screen.
【저자키워드】 SARS-CoV-2, Restriction factors, 【초록키워드】 public health, ACE2, Coronaviruses, Transmission, virus, angiotensin-converting enzyme 2, Spread, CRISPR, SARS-CoV-2 spike protein, virus entry, expression, Critical, Factor, subunit, residue, S1/S2, cleavage site, insertion, plasma membrane, clone, S1/S2 boundary, Host, Affect, Cell, highlight, S1/S2 site, endosomal entry pathway, identify, reduced, affecting, disrupting, entry of SARS-CoV-2, genome-wide CRISPR screen, modulating, multi-basic cleavage site, 【제목키워드】 regulate, Factor, SARS-CoV-2 entry, Host, identify, genome-wide CRISPR screen,