Comprehensive Functional Analysis of N-Linked Glycans on Ebola Virus GP1

Lennemann, Nicholas J.; Rhein, Bethany A.; Ndungo, Esther; Chandran, Kartik; Qiu, Xiangguo; Maury, Wendy

Domain-specific N-linked deglycosylation alters C-type lectin utilization. CLECs, which mediate GP-dependent entry, are expressed on a wide variety of EBOV target cell types, including macrophages, dendritic cells, and hepatic cells, and each binds to specific glycan moieties (25) (26) (27) (28) (29) . To better understand the role of EBOV GP1 N-linked glycans in CLEC-dependent tropism, we determined the receptor utilization of mutants that lack N-linked glycans on the GP1 core (7G), the MLD (GPm8G), or both (7Gm8G). Constructs expressing Myc-tagged CLECs were transfected into poorly permissive HEK293T cells. To enhance expression, the ectodomains of CLECs DC-SIGN and human MGL (hMGL) were fused to the transmembrane and cytoplasmic domain of L-SIGN (resulting in constructs L DC-SIGN and L hMGL, respectively). All constructs expressed well in transfected cells (Fig. S5) .