Structure, function and evolution of the trypanosome nuclear envelope . (360G-Wellcome-082813_Z_07_Z)

We seek a molecular characterisation of the trypanosome nuclear pore complex (NPC) and envelope. Our rational for initiating this project includes the divergent evolutionary position of trypanosomes, unique aspects of mRNA maturation (polycistronic transcription), unusual aspects of control of mRNA stability (primarily via elements within the 3 UTR) and a clear absence of a conventional (metazoan-related) nuclear lamina. We aim to: 1. Produce a high-resolution connectivity map of the trypano some NPC for definition of both composition and architecture. These data will facilitate a complete reconstruction of the trypanosome NPC for evolutionary and functional comparisons with yeast and mammalian NPCs. 2. Assess the functions of individual trypanosome nuclear pore proteins in terms of viability, structure and interactions with the nuclear lamina. Again comparison with studies in yeast and mammals will provide a framework for definition of divergence and conservation of function of sp ecific elements. 3. Initiate analysis of the trypanosome nuclear lamina. Currently this is unexplored, and our investigations have identified several strong candidates as components of this structure. All three aims will exploit a recently completed characterisation of the Trypanosoma brucei nuclear envelope proteome performed in collaboration between the applicants and the Chait laboratories (supporting data).