FASN inhibitor TVB-3166 prevents S-acylation of the spike protein of human coronaviruses
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 sp...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-09-01
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| Series: | Journal of Lipid Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S002222752200089X |
| _version_ | 1811264402812305408 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Katrina Mekhail |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-12T20:03:46Z |
| format | Article |
| id | doaj.art-8b95179bea8a414e9cff10cdfb16ac25 |
| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-04-12T20:03:46Z |
| publishDate | 2022-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-8b95179bea8a414e9cff10cdfb16ac252022-12-22T03:18:28ZengElsevierJournal of Lipid Research0022-22752022-09-01639100256FASN inhibitor TVB-3166 prevents S-acylation of the spike protein of human coronavirusesKatrina Mekhail0Minhyoung Lee1Michael Sugiyama2Audrey Astori3Jonathan St-Germain4Elyse Latreille5Negar Khosraviani6Kuiru Wei7Zhijie Li8James Rini9Warren L. Lee10Costin Antonescu11Brian Raught12Gregory D. Fairn13Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada; Keenan Research Centre, St. Michael’s Hospital, Unity Health Toronto, Toronto, Ontario, CanadaDepartment of Biochemistry, University of Toronto, Toronto, Ontario, Canada; Keenan Research Centre, St. Michael’s Hospital, Unity Health Toronto, Toronto, Ontario, CanadaDepartment of Chemistry and Biology, Ryerson University, Toronto, Ontario, CanadaPrincess Margaret Cancer Centre, University Health Network, Ontario, CanadaPrincess Margaret Cancer Centre, University Health Network, Ontario, CanadaKeenan Research Centre, St. Michael’s Hospital, Unity Health Toronto, Toronto, Ontario, Canada; Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario, CanadaKeenan Research Centre, St. Michael’s Hospital, Unity Health Toronto, Toronto, Ontario, CanadaKeenan Research Centre, St. Michael’s Hospital, Unity Health Toronto, Toronto, Ontario, CanadaDepartment of Biochemistry, University of Toronto, Toronto, Ontario, CanadaDepartment of Biochemistry, University of Toronto, Toronto, Ontario, Canada; Department of Molecular Genetics, University of Toronto, Ontario, CanadaDepartment of Biochemistry, University of Toronto, Toronto, Ontario, Canada; Keenan Research Centre, St. Michael’s Hospital, Unity Health Toronto, Toronto, Ontario, Canada; Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario, Canada; Department of Medicine, University of Toronto, Toronto, Ontario, CanadaDepartment of Chemistry and Biology, Ryerson University, Toronto, Ontario, CanadaPrincess Margaret Cancer Centre, University Health Network, Ontario, Canada; Department of Medical Biophysics, University of Toronto, Ontario, CanadaDepartment of Biochemistry, University of Toronto, Toronto, Ontario, Canada; Keenan Research Centre, St. Michael’s Hospital, Unity Health Toronto, Toronto, Ontario, Canada; Department of Surgery, University of Toronto, Toronto, Ontario, Canada; Department of Pathology, Dalhousie University, Halifax, Nova Scotia, Canada; For correspondence: Gregory D. FairnThe 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.http://www.sciencedirect.com/science/article/pii/S002222752200089XS-acylationZDHHCspike coronavirusclick chemistryFASNpalmitate |
| spellingShingle | 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 FASN inhibitor TVB-3166 prevents S-acylation of the spike protein of human coronaviruses Journal of Lipid Research S-acylation ZDHHC spike coronavirus click chemistry FASN palmitate |
| title | FASN inhibitor TVB-3166 prevents S-acylation of the spike protein of human coronaviruses |
| title_full | FASN inhibitor TVB-3166 prevents S-acylation of the spike protein of human coronaviruses |
| title_fullStr | FASN inhibitor TVB-3166 prevents S-acylation of the spike protein of human coronaviruses |
| title_full_unstemmed | FASN inhibitor TVB-3166 prevents S-acylation of the spike protein of human coronaviruses |
| title_short | FASN inhibitor TVB-3166 prevents S-acylation of the spike protein of human coronaviruses |
| title_sort | fasn inhibitor tvb 3166 prevents s acylation of the spike protein of human coronaviruses |
| topic | S-acylation ZDHHC spike coronavirus click chemistry FASN palmitate |
| url | http://www.sciencedirect.com/science/article/pii/S002222752200089X |
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