Here we devised a chemoenzymatic labeling strategy to site-specifically append probes to the influenza hemagglutinin (HA) and neuraminidase (NA) proteins using the bacterial sortase A enzyme. Because labeling is confined to surface exposed HA and NA in the context of live, infected cells, it is possible to study budding biochemically and microscopically in real-time. Using this system, we can observe budding of flu virions from discrete sites at the cell surface. Our work will enable detailed investigation into the birth of viruses from infected host cells and can likely be applied to viruses other than influenza that have been similarly resistant to real-time microscopic observation during budding.
Virology tools just get better and better.