The sensor of VSG coat integrity on the surface of Trypanosoma brucei (360G-Wellcome-105405_Z_14_Z)

Trypanosoma brucei, the causative agent of Human African Trypanosomiasis, constantly changes its variant surface glycoprotein (VSG) coat to avoid elimination by the immune system of the mammalian host. It is known that VSG synthesis is essential: RNAi knockdown of VSG results in precytokinesis arrest. However, it is not yet known how the cell senses that the VSG coat is intact before cell division occurs. This project seeks to identify the molecular mechanism of the sensor of VSG coat integrity. I will replace wild-type VSG with a number of different VSG mutants. These include a truncated form without a GPI anchor and a VSG with a trans membrane anchor. I will assess whether these cells are capable of division and compare the growth rates to cell lines expressing wild-type VSG. I will also investigate the requirements for VSG recycling by determining whether a VSG-GFP fusion mutant is recycled on the cell surface. To identify the genes responsible for controlling surface VSG density I will carry out a forward genetic screen using RITSeq. The identification of the VSG density sensing signal pathway will be novel biology and inform future drug design.