A systems approach for understanding cell surface dynamics in trypanosomes (360G-Wellcome-204697_Z_16_Z)

The surface membrane and endosomal system of protozoan pathogens directly interfaces with the host; mediated functions including immune evasion, environmental motoring and drug interactions. Recently it has emerged that surface composition and the mechanisms controlling membrane protein turnover in trypanosomatids are highly divergent, and that these processes also are connected with drug interactions. We have clear evidence for complex control based around ubiquitylation, but the roles of many surface proteins remain cryptic. I propose a program of work to integrate, in a system wide manner, how these novel aspects of biology converge to maintain the parasite surface. 1. Examine the mechanisms of ISG and AQP turnover,2. Evaluate the potential of ISGs for novel anti-trypanosome drug delivery,3. Assess the essentiality of ISG protein families in vitro and in vivo using genome editing.4. Refine our understanding of the clathrin and ESCRT sorting systems of trypanosomes to determine how these processes mediate the transport of ISGs, AQPs and the surface proteome in general. Each of these aspects will exploit a combination of state of the art proteomics, super-resolution microscopy and novel genetic tools to understand in greater detail the importance of surface proteins to trypanosome biology, virulence and infectivity.