FASN inhibitor TVB-3166 prevents S-acylation of the spike protein of human coronaviruses |
| |
Authors: | Katrina Mekhail Minhyoung Lee Michael Sugiyama Audrey Astori Jonathan St-Germain Elyse Latreille Negar Khosraviani Kuiru Wei Zhijie Li James Rini Warren L. Lee Costin Antonescu Brian Raught Gregory D. Fairn |
| |
Affiliation: | 1. Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada;2. Keenan Research Centre, St. Michael’s Hospital, Unity Health Toronto, Toronto, Ontario, Canada;3. Department of Chemistry and Biology, Ryerson University, Toronto, Ontario, Canada;4. Princess Margaret Cancer Centre, University Health Network, Ontario, Canada;5. Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario, Canada;6. Department of Molecular Genetics, University of Toronto, Ontario, Canada;7. Department of Medicine, University of Toronto, Toronto, Ontario, Canada;8. Department of Medical Biophysics, University of Toronto, Ontario, Canada;9. Department of Surgery, University of Toronto, Toronto, Ontario, Canada;10. Department of Pathology, Dalhousie University, Halifax, Nova Scotia, Canada |
| |
Abstract: | The spike protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and other coronaviruses mediates host cell entry and is S-acylated on multiple phylogenetically conserved cysteine residues. Multiple protein acyltransferase enzymes have been reported to post-translationally modify spike proteins; however, strategies to exploit this modification are lacking. Using resin-assisted capture MS, we demonstrate that the spike protein is S-acylated in SARS-CoV-2-infected human and monkey epithelial cells. We further show that increased abundance of the acyltransferase ZDHHC5 associates with increased S-acylation of the spike protein, whereas ZDHHC5 knockout cells had a 40% reduction in the incorporation of an alkynyl-palmitate using click chemistry detection. We also found that the S-acylation of the spike protein is not limited to palmitate, as clickable versions of myristate and stearate were also labelled the protein. Yet, we observed that ZDHHC5 was only modified when incubated with alkyne-palmitate, suggesting it has specificity for this acyl-CoA, and that other ZDHHC enzymes may use additional fatty acids to modify the spike protein. Since multiple ZDHHC isoforms may modify the spike protein, we also examined the ability of the FASN inhibitor TVB-3166 to prevent S-acylation of the spike proteins of SARS-CoV-2 and human CoV-229E. We show that treating cells with TVB-3166 inhibited S-acylation of expressed spike proteins and attenuated the ability of SARS-CoV-2 and human CoV-229E to spread in vitro. Our findings further substantiate the necessity of CoV spike protein S-acylation and demonstrate that de novo fatty acid synthesis is critical for the proper S-acylation of the spike protein. |
| |
Keywords: | S-acylation ZDHHC spike coronavirus click chemistry FASN palmitate SARS-CoV-2 CoV-229E host cell entry infection ACE2" },{" #name" :" keyword" ," $" :{" id" :" kwrd0065" }," $$" :[{" #name" :" text" ," _" :" angiotensin-converting enzyme 2 acyl-RAC" },{" #name" :" keyword" ," $" :{" id" :" kwrd0075" }," $$" :[{" #name" :" text" ," _" :" acyl-resin-assisted capture APE" },{" #name" :" keyword" ," $" :{" id" :" kwrd0085" }," $$" :[{" #name" :" text" ," $$" :[{" #name" :" __text__" ," _" :" acyl-polyethylene glycol " },{" #name" :" underline" ," _" :" e" },{" #name" :" __text__" ," _" :" xchange CoV" },{" #name" :" keyword" ," $" :{" id" :" kwrd0095" }," $$" :[{" #name" :" text" ," _" :" coronavirus COVID-19" },{" #name" :" keyword" ," $" :{" id" :" kwrd0105" }," $$" :[{" #name" :" text" ," _" :" coronavirus disease 2019 Cy5.5" },{" #name" :" keyword" ," $" :{" id" :" kwrd0115" }," $$" :[{" #name" :" text" ," _" :" cyanine 5.5 Cys" },{" #name" :" keyword" ," $" :{" id" :" kwrd0125" }," $$" :[{" #name" :" text" ," _" :" cysteine DAPI" },{" #name" :" keyword" ," $" :{" id" :" kwrd0135" }," $$" :[{" #name" :" text" ," _" :" 4',6-diamidino-2-phenylindole D-PBS" },{" #name" :" keyword" ," $" :{" id" :" kwrd0145" }," $$" :[{" #name" :" text" ," _" :" Dulbecco’s PBS DSP" },{" #name" :" keyword" ," $" :{" id" :" kwrd0155" }," $$" :[{" #name" :" text" ," _" :" dithiobissuccinimidyl propionate EMEM" },{" #name" :" keyword" ," $" :{" id" :" kwrd0165" }," $$" :[{" #name" :" text" ," _" :" Eagle's MEM HA" },{" #name" :" keyword" ," $" :{" id" :" kwrd0175" }," $$" :[{" #name" :" text" ," _" :" hemagglutinin 15-HDYA" },{" #name" :" keyword" ," $" :{" id" :" kwrd0185" }," $$" :[{" #name" :" text" ," _" :" 15-hexadecynoic acid HEK293T" },{" #name" :" keyword" ," $" :{" id" :" kwrd0195" }," $$" :[{" #name" :" text" ," _" :" human embryonic kidney 293T cell line MHV" },{" #name" :" keyword" ," $" :{" id" :" kwrd0205" }," $$" :[{" #name" :" text" ," _" :" murine hepatitis virus MOI" },{" #name" :" keyword" ," $" :{" id" :" kwrd0215" }," $$" :[{" #name" :" text" ," _" :" multiplicity of infection NEM" },{" #name" :" keyword" ," $" :{" id" :" kwrd0225" }," $$" :[{" #name" :" text" ," $$" :[{" #name" :" italic" ," _" :" N" },{" #name" :" __text__" ," _" :" -ethylmaleimide PEG" },{" #name" :" keyword" ," $" :{" id" :" kwrd0235" }," $$" :[{" #name" :" text" ," _" :" polyethylene glycol PFA" },{" #name" :" keyword" ," $" :{" id" :" kwrd0245" }," $$" :[{" #name" :" text" ," _" :" paraformaldehyde SARS-CoV-2" },{" #name" :" keyword" ," $" :{" id" :" kwrd0255" }," $$" :[{" #name" :" text" ," _" :" severe acute respiratory syndrome coronavirus 2 ZDHHC" },{" #name" :" keyword" ," $" :{" id" :" kwrd0265" }," $$" :[{" #name" :" text" ," _" :" zinc finger Asp-His-His-Cys |
本文献已被 ScienceDirect 等数据库收录! |
|