TY - JOUR
T1 - Photochemical Identification of Auxiliary Severe Acute Respiratory Syndrome Coronavirus 2 Host Entry Factors Using μmap
AU - Suzuki, Saori
AU - Geri, Jacob B.
AU - Knutson, Steve D.
AU - Bell-Temin, Harris
AU - Tamura, Tomokazu
AU - Fernández, David F.
AU - Lovett, Gabrielle H.
AU - Till, Nicholas A.
AU - Heller, Brigitte L.
AU - Guo, Jinchao
AU - Macmillan, David W.C.
AU - Ploss, Alexander
N1 - Funding Information:
This work was supported by kind gifts from Merck, BMS, Pfizer, Janssen, Genentech, and Eli Lilly. We also acknowledge the Princeton Catalysis Initiative for supporting this work. The authors thank Saw Kyin and Henry H. Shwe at the Princeton Proteomics Facility. We also thank Mohsan Saeed (Boston University) for providing A549-ACE2/TMPRSS2 cells,pLOC-ACE2-PuroR, and SARS-CoV2-S_B.1.1.529_codon optimized plasmid and Celeste Nelson for providing the Calu-3, BEAS-2B, and NuLi-1 cell lines. We are grateful to Christina DeCoste and Katherine Rittenbach of the Flow Cytometry Resource Facility at Princeton University for their assistance in the planning and execution of all flow cytometry experiments. We would also like to thank Gary S. Laevsky and Sha Wang of the Confocal Imaging Facility, a Nikon Center of Excellence, in the Department of Molecular Biology at Princeton University for instrument use and technical advice.
Funding Information:
This study was funded in part by grants from the National Institutes of Health (R01AI138797, R01AI107301, R01AI146917, and R01AI153236 to A.P.), a Burroughs Wellcome Fund Award for Investigators in Pathogenesis (101539 to A.P.), and Princeton COVID-19 research funds through the Office of the Dean for Research and a component of the National Institutes of Health (NIH) under award number UL1TR003017 (to A.P.). This work was also funded by the NIH National Institute of General Medical Sciences (R35-GM134897-02). S.D.K. acknowledges the NIH for a postdoctoral fellowship (1F32GM142206-01). The Molecular Biology Flow Cytometry Resource Facility is partially supported by the Cancer Institute of New Jersey Cancer Center Support grant (P30CA072720). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
Publisher Copyright:
© 2022 American Chemical Society. All rights reserved.
PY - 2022/9/14
Y1 - 2022/9/14
N2 - Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the infectious agent of the COVID-19 pandemic, remains a global medical problem. Angiotensin-converting enzyme 2 (ACE2) was identified as the primary viral entry receptor, and transmembrane serine protease 2 primes the spike protein for membrane fusion. However, ACE2 expression is generally low and variable across tissues, suggesting that auxiliary receptors facilitate viral entry. Identifying these factors is critical for understanding SARS-Cov-2 pathophysiology and developing new countermeasures. However, profiling host-virus interactomes involves extensive genetic screening or complex computational predictions. Here, we leverage the photocatalytic proximity labeling platform μMap to rapidly profile the spike interactome in human cells and identify eight novel candidate receptors. We systemically validate their functionality in SARS-CoV-2 pseudoviral uptake assays with both Wuhan and Delta spike variants and show that dual expression of ACE2 with either neuropilin-2, ephrin receptor A7, solute carrier family 6 member 15, or myelin and lymphocyte protein 2 significantly enhances viral uptake. Collectively, our data show that SARS-CoV-2 synergistically engages several host factors for cell entry and establishes μMap as a powerful tool for rapidly interrogating host-virus interactomes.
AB - Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the infectious agent of the COVID-19 pandemic, remains a global medical problem. Angiotensin-converting enzyme 2 (ACE2) was identified as the primary viral entry receptor, and transmembrane serine protease 2 primes the spike protein for membrane fusion. However, ACE2 expression is generally low and variable across tissues, suggesting that auxiliary receptors facilitate viral entry. Identifying these factors is critical for understanding SARS-Cov-2 pathophysiology and developing new countermeasures. However, profiling host-virus interactomes involves extensive genetic screening or complex computational predictions. Here, we leverage the photocatalytic proximity labeling platform μMap to rapidly profile the spike interactome in human cells and identify eight novel candidate receptors. We systemically validate their functionality in SARS-CoV-2 pseudoviral uptake assays with both Wuhan and Delta spike variants and show that dual expression of ACE2 with either neuropilin-2, ephrin receptor A7, solute carrier family 6 member 15, or myelin and lymphocyte protein 2 significantly enhances viral uptake. Collectively, our data show that SARS-CoV-2 synergistically engages several host factors for cell entry and establishes μMap as a powerful tool for rapidly interrogating host-virus interactomes.
UR - http://www.scopus.com/inward/record.url?scp=85137864921&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85137864921&partnerID=8YFLogxK
U2 - 10.1021/jacs.2c06806
DO - 10.1021/jacs.2c06806
M3 - Article
C2 - 36049228
AN - SCOPUS:85137864921
SN - 0002-7863
VL - 144
SP - 16604
EP - 16611
JO - Journal of the American Chemical Society
JF - Journal of the American Chemical Society
IS - 36
ER -