Structural and functional analysis of interactions involving the nuclear transport machinery that generate nuclear RNA and protein export and orchestrate mitotic spindle assembly. (360G-Wellcome-080522_Z_06_Z)

My overall goal is to understand how nuclear transport factors interact with their cargoes and other components of the transport machinery to (i) export macromolecules from the nucleus during interphase and (ii) orchestrate spindleassembly during mitosis. These aims share a common focus on nuclear transportfactors and their interaction with different partners, and build on my previous work that established the structural basis for key steps in the nuclear protein import cycle. Molecular recognition by transport factors is fundamental to the nuclear export of proteins, mRNA, pre-microRNA, and tRNA, and is also crucial in mitotic spindle organization. My primary goal is to usethe interdisciplinary approach I developed for studying nuclear protein import, in which structural, biochemical, molecular and cellular methods will be used in concert to investigate the key complexes involved in each process and establish interaction interfaces. This information will then be used to define how each function is generated by a precisely orchestrated series of interactions, employing structure-based engineered mutants that alter specificinteractions. These mutants will be used to establish the spatial and temporalpattern of interactions that generate function and so enable these important cellular processes to be understood at the molecular level.