FASN inhibitor TVB-3166 prevents S-acylation of the spike protein of human coronaviruses

Mekhail, Katrina; Lee, Minhyoung; Sugiyama, Michael G.; Astori, Audrey; St-Germain, Jonathan; Latreille, Elyse; Khosraviani, Negar; Wei, Kuiru; Li, Zhijie; Rini, James M.; Lee, Warren L.; Antonescu, Costin N.; Raught, Brian; Fairn, Gregory D.

doi:10.1016/j.jlr.2022.100256

Cited by 6 publications

(6 citation statements)

References 82 publications

(123 reference statements)

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“…At this stage, though, we cannot exclude that the N-terminal extension additionally improves the enzymatic activity and/or substrate recognition of ZDHHC20 Long . Spike S-acylation upon transfection into culture cells has been monitored by many laboratories (3,4,6,7,29,30), yet we all overlooked the fact that ZDHHC20 Short , which we now know was the main enzyme expressed in these cells, is very inefficient in modifying Spike. This is likely because 3 H-palmitic acid incorporation and acyl-capture assays monitor gains of signal such that abundant proteins with numerous cysteines, like over-expressed Spike, will produce readily detectable signals suggestive of extensive acylation.…”

Section: Zdhhc20 Long Has Greatly Enhanced Performance In Modifying S...mentioning

confidence: 99%

“…However, these methods do not inform on the stoichiometry of S-acylation. PEGylation experiments have been done, but the minimal band shifts of Spike that were observed were not interpreted (4,7,29,31), yet did still indicate that acylation under these conditions was occurring on very few Spike molecules within the population. Instead, Spike only becomes extensively acylated during infection when cells express significant levels of ZDHHC20 Long .…”

Section: Zdhhc20 Long Has Greatly Enhanced Performance In Modifying S...mentioning

confidence: 99%

See 1 more Smart Citation

SARS-CoV-2 shifts transcription of host gene to increase Spike acylation and boost infectivity

Mesquita

Abrami

Bracq

et al. 2023

Preprint

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SARS-CoV-2 infection requires Spike protein mediating fusion between the viral and cellular membranes. The fusogenic activity of Spike requires its post-translational lipid modification by host S-acyltransferases, predominantly ZDHHC20. Previous observations indicate that SARS-CoV-2 infection augments the S-acylation of Spike when compared to transfection. Here, we find that SARS-CoV-2 infection triggers a change in the transcriptional start site of the zddhc20 gene, both in cells and in an in vivo infection model, resulting in a 67-amino-acid-long N-terminally extended protein with 37-times higher Spike acylating activity, leading to enhanced viral infectivity. Furthermore, we observed the same induced transcriptional change in response to other challenges, such as chemically induced colitis, indicating that SARS-CoV-2 hijacks an existing cell damage response pathway to generate more infectious viruses.

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Section: Zdhhc20 Long Has Greatly Enhanced Performance In Modifying S...mentioning

confidence: 99%

Section: Zdhhc20 Long Has Greatly Enhanced Performance In Modifying S...mentioning

confidence: 99%

SARS-CoV-2 shifts transcription of host gene to increase Spike acylation and boost infectivity

Mesquita

Abrami

Bracq

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

“…Examples include membrane trafficking of proto-oncogene Ras 5,6 and Src 7,8 isoforms, badrenergic receptor signaling [9][10][11] , vesicle/vacuole fusion 12,13 and proper assembly of respiratory viruses [14][15][16] . Large scale computational analyses suggest that aberrant S-palmitoylation may play a role in cancer and neurologic disorders 17 and pharmacological studies targeting the S-palmitoylation machinery show promise for the future development of antiviral 18,19 and antineoplastic 20,21 therapies.…”

Section: Introductionmentioning

confidence: 99%

Analysis of Protein Cysteine Acylation Using a Modified Suspension Trap (Acyl-Trap)

Forrester,

Egol,

Tata

et al. 2024

Preprint

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Protein cysteine thiols undergo reversibleS-acylation via a thioester linkage in vivo.S-palmitoylation, modification by C16:0 fatty acid, is a commonS-acylation that mediates protein-membrane and protein-protein interactions critical to an array of biological processes, from homeostatic lung surfactant function to cellular transformation. The most widely usedS-acylation assays, including acyl-biotin exchange (ABE) and acyl resin-assisted capture (Acyl-RAC), utilize blocking of free Cys thiols, hydroxylamine-dependent cleavage of the thioester and subsequent labeling of nascent thiol. ABE and Acyl-RAC have enabled both the proteome-wide identification ofS-palmitoylation sites and basic biochemical studies. Yet, these assays generally utilize hundreds of micrograms to milligrams of input material and require numerous reagent removal and washing steps, making them laborious and ill-suited for high throughput and low input applications. To overcome this, we devisedAcyl-Trap, a suspension trap-based assay that utilizes a thiol-reactive quartz to enable buffer exchange and hydroxylamine-mediatedS-acyl enrichment from 20-50 micrograms of input protein. The method is compatible with protein-level detection ofS-acylated proteins as well asS-acyl site-based identification and quantification using on-quartz isobaric (tandem mass tag) labeling and LC-MS/MS. Also described are conditions for long-term hydroxylamine storage, which further expedites the assay and minimizes waste. More generally, Acyl-Trap serves as a proof-of-concept for PTM-tailored suspension traps suitable for both traditional intact protein detection and chemoproteomic workflows.

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“…At this stage, though, we cannot exclude that the N-terminal extension additionally improves the enzymatic activity and/or substrate recognition of ZDHHC20 Long . Spike S-acylation upon transfection into culture cells has been monitored by many laboratories 3,4,6,7,29,30 , yet we all overlooked the fact that ZDHHC20 Short , which we now know was the main enzyme expressed in these cells, is very ine cient in modifying Spike. This is likely because 3 H-palmitic acid incorporation and acyl-capture assays monitor gains of signal such that abundant proteins with numerous cysteines, like over-expressed Spike, will produce readily detectable signals suggestive of extensive acylation.…”

mentioning

confidence: 99%

“…However, these methods do not inform on the stoichiometry of S-acylation. PEGylation experiments have been done, but the minimal band shifts of Spike that were observed were not interpreted 4,7,29,31 , yet did still indicate that acylation under these conditions was occurring on very few Spike molecules within the population. Instead, Spike only becomes extensively acylated during infection when cells express signi cant levels of ZDHHC20 Long .…”

mentioning

confidence: 99%

SARS-CoV-2 shifts transcription of host gene to increase Spike acylation and boost infectivity

Goot¹,

Mesquita²,

Abrami

et al. 2023

Preprint

View full text Add to dashboard Cite

SARS-CoV-2 infection requires Spike protein mediating fusion between the viral and cellular membranes. The fusogenic activity of Spike requires its post-translational lipid modification by host S-acyltransferases, predominantly ZDHHC20. Previous observations indicate that SARS-CoV-2 infection augments the S-acylation of Spike when compared to transfection. Here, we find that SARS-CoV-2 infection triggers a change in the transcriptional start site of the zddhc20 gene, both in cells and in an in vivo infection model, resulting in a 67-amino–acid-long N-terminally extended protein with 37-times higher Spike acylating activity, leading to enhanced viral infectivity. Furthermore, we observed the same induced transcriptional change in response to other challenges, such as chemically induced colitis, indicating that SARS-CoV-2 hijacks an existing cell damage response pathway to generate more infectious viruses.

show abstract

FASN inhibitor TVB-3166 prevents S-acylation of the spike protein of human coronaviruses

Cited by 6 publications

References 82 publications

SARS-CoV-2 shifts transcription of host gene to increase Spike acylation and boost infectivity

SARS-CoV-2 shifts transcription of host gene to increase Spike acylation and boost infectivity

Analysis of Protein Cysteine Acylation Using a Modified Suspension Trap (Acyl-Trap)

SARS-CoV-2 shifts transcription of host gene to increase Spike acylation and boost infectivity

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