FUT8-mediated core fucosylation of receptor APN drives entry of multiple alphacoronaviruses

Understanding the interaction mechanisms between coronaviruses (CoVs) and their hosts is crucial for understanding the viral replication cycle and identifying novel antiviral targets. In this study, we found alpha-(1,6)-fucosyltransferase (FUT8), via its fucosyltransferase activity, is involved in the several alphacoronaviruses (α-CoVs) spike-receptor aminopeptidase N (APN) interaction to regulate viral entry. Mechanistically, pAPN lacking FUT8-mediated modification showed no binding to the transmissible gastroenteritis virus (TGEV) RBD. The viral entry depends on core fucosylation at pAPN N736. Further pAPN glycoproteomic analysis confirmed that core fucosylation at N736 is indeed present in wild-type (WT) cells but almost abolished in KO cells, highlighting that FUT8 facilitates viral entry by mediating core fucosylation of pAPN N736. Interestingly, FUT8 is also essential for the entry of canine and feline CoVs, which use APN as their receptor, by mediating core fucosylation at N747 and N740 on canine and feline APN, respectively, demonstrating that FUT8 has a conserved function across these species. Overall, this study uncovers the role of FUT8 in multiple α-CoVs entry, revealing the importance of core fucosylation in viral replication and identifying FUT8 as a potential broad-spectrum antiviral target.