Exploring Filovirus-host interactions at single cell resolution

Enveloped viruses are commonly coated with highly glycosylated glycoproteins termed "spikes", that offer a degree of multivalency through trimeric organisation. Similarly, the cell surface, as well as organelle membranes, are enriched in glycoproteins, importantly being the first site of contact for an invading virus. Glycans not only play a role in the initial interaction between virus and host, but are of central importance for immunological recognition through the masking of epitopes under dense "glycan shields". However, membrane environments and the inherent flexibility of individual glycans hinders the use of traditional ensemble-based structural techniques to determine glycan specificity in mediating interactions. This project will establish an entirely new way to study glycoprotein interactions in cellular environments with unprecedented spatiotemporal resolution. Harnessing metabolic glycoprotein engineering, we will develop cross-linking mass spectrometry to selectively attach glycans or glycoproteins with their respective interaction partners at a single cell resolution. Utilising state- of-the-art developments in timsTOF-based mass spectrometry, alongside single- cell proteomics, we will track Filovirus cellular interactions throughout the viral lifecycle with atomic resolution. By gaining insights into Filovirus interactions, we hope to improve our understanding of viral evolution and host adaptation with the overarching goal to aid in the development of antiviral and therapeutic approaches.