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.