Аннотация:Influenza A viruses contain two S‐acylated proteins, the ion channel M2 and the glycoprotein hemagglutinin (HA). Acylation of the latter is essential for virus replication. Here we analyzed expression of each of the 23 members of the family of ZDHHC acyltransferases in human airway cells, the site of virus replication. RT‐PCR revealed that every ZDHHC acyltransferase (except ZDHHC19) is expressed in A549 and Calu cells. Interestingly, expression of one ZDHHC, ZDHHC22, is upregulated in virus‐infected cells; this effect is more pronounced after infection with an avian compared to a human virus strain. The viral protein NS1 triggers ZDHHC22 expression in transfected cells, whereas recombinant viruses lacking a functional NS1 gene did not cause ZDHHC22 upregulation. CRISPR/Cas9 technology was then used to knock‐out the ZDHHC22 gene in A549 cells. However, acylation of M2 and HA was not reduced, as analyzed for intracellular HA and M2 and the stoichiometry of S‐acylation of HA incorporated into virus particles did not change according to MALDI‐TOF mass spectrometry analysis. Comparative mass spectrometry of palmitoylated proteins in wt and ΔZDHHC22 cells identified 25 potential substrates of ZDHHC22 which might be involved in virus replication.
