Functional assessment of cell entry and receptor usage for lineage B β-coronaviruses, including 2019-nCoV. (2020) Michael Letko and Vincent Munster. bioRxiv doi: https://doi.org/10.1101/2020.01.22.915660
We will adhere to the Universal Principled (UP) Review guidelines proposed in:
Universal Principled Review: A Community-Driven Method to Improve Peer Review.Krummel M, Blish C, Kuhns M, Cadwell K, Oberst A, Goldrath A, Ansel KM, Chi H, O’Connell R, Wherry EJ, Pepper M; Future Immunology Consortium.Cell . 2019 Dec 12;179(7):1441-1445. doi: 10.1016/j.cell .2019.11.029.
SUMMARY: Letko and Munster report a new functional viromics platform whereby receptor binding domains (RBDs) from different lineage B betacoronaviruses were cloned into a codon-optimized gene for SARS CoV spike protein which was then incorporated into pseudotyped VSV particles for functional assays. Entry was indicated by luciferase reporter activity. This screen facilitated rapid identification of RBD-receptor interactions, with less expense than previous methods. The authors confirmed previous findings that only RBDs belonging to clade-1 of the B-lineage of beta-coronaviruses use the ACE2 receptor. Furthermore, the authors showed that for a variety of B-lineage coronaviruses, protease treatment prior to infection enhances entry into different cell types from different species. They confirmed that protease treatment enhanced receptor-dependent viral entry. By introducing 14 amino acids known to contact the ACE2 receptor into clade-2 and clade-3 RBDs, the authors confirmed that these AAs are important for ACE2 recognition. They also determined that the surrounding AA sequence context is important for ACE2 recognition. Finally, they showed that the new 2019-nCoV coronavirus (now known as SARS-CoV-2 according to the ICTV) is related to Clade-1 betacoronaviruses, similar to SARS, and also utilizes the ACE2 entry receptor.