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

The spike protein of severe acute respiratory system syndrome coronavirus 2 (SARS-CoV-2) along with other coronaviruses mediates host cell entry and it is S-acylated on multiple phylogenetically conserved cysteine residues. Multiple protein acyltransferase enzymes happen to be reported to publish-translationally modify spike proteins however, ways of exploit this modification are missing. Using resin-aided capture MS, we show the spike proteins are S-acylated in SARS-CoV-2-infected human and monkey epithelial cells. We further reveal that elevated abundance from the acyltransferase ZDHHC5 associates with elevated S-acylation from the spike protein, whereas ZDHHC5 knockout cells were built with a 40% decrease in the incorporation of the alkynyl-palmitate using click chemistry recognition. We discovered that the S-acylation from the spike proteins are not restricted to palmitate, as clickable versions of myristate and stearate were also labelled the protein. Yet, we observed that ZDHHC5 was just modified when incubated with alkyne-palmitate, suggesting it’s specificity with this acyl-CoA, which other ZDHHC enzymes could use additional essential fatty acids to change the spike protein. Since multiple ZDHHC isoforms may customize the spike protein, we examined ale the FASN inhibitor TVB-3166 to avoid S-acylation from the spike proteins of SARS-CoV-2 and human CoV-229E. We reveal that treating cells with TVB-3166 inhibited S-acylation of expressed spike proteins and attenuated ale SARS-CoV-2 and human CoV-229E to spread in vitro. Our findings further substantiate involve CoV spike protein S-acylation and show de novo essential fatty acid synthesis is crucial for that proper S-acylation from the spike protein.