Structure and function of the mammalian cell nucleus (360G-Wellcome-073980_Z_03_B)

My research programme is aimed at understanding how the nucleus is organised in mammalian cells, how specific subnuclear bodies are assembled and regulated and how nuclear structure is related to biological function and to human molecular disease. A major theme of the programme is to use a combination of quantitative in vivo and in vitro techniques to measure dynamic processes in the cell nucleus. To achieve these aims we will focus primarily on studying the dynamic properties of the nucleolus and the nuclear organisation and function of the pre-mRNA splicing machinery. We will also study the role of reversible protein phosphorylation in regulating both nucleolar interactions and the splicing mechanism. We will compare data derived from biochemical experiments carried out using nuclear extracts prepared from cultured cell lines with data on the movement and interactions of proteins in vivo in the same cell lines. We will use proteomic methods to analyse and quantitate the protein composition of nucleoli under different growth and metabolic conditions and we will characterise dynamic interactions in live cells by imaging Fluorescence Protein-tagged fusion proteins. In parallel, we will analyse the organisation and structure of nuclear factors in both transformed and normal human tissue, derived from patient biopsies, to maximise the opportunities for our work on the cell biology of the nucleus to enhance clinical practice.