Research objectives and content
I shall analyse the actin binding properties, cellular distribution and function of ADF and cofilin, essential proteins involved in cytoskeletal remodelling, e.g. in cytokinesis and locomotion. Dr Weeds's laboratory and I are each well equipped to perform this work together. We have specific antibodies for ADF, cofilin and nuclear actin raised from three different animal sources. Preparing GFP-tagged proteins will permit us to investigate: (i) cellular distribution, response to stimuli and the role of two such similar proteins in one cell (ii) the role of phosphorylation and its regulation on intracellular localization and function of ADF/cofilin and (iii) the translocation of actin and cofilin to the nucleus. Using GFP-tags will enable us to monitor these effects in real time. We shall then be able to correlate biochemical evidence with in vivo studies which neither of us are equipped to do in isolation. Training Content
I have the cell biology and immunofluorescence expertise required by Dr Weeds and his group to perform in vivo analyses and in return they will instruct me further in biochemistry and the necessary molecular biology skills to prepare transfection, expression and mutated cofilin/ADF constructs.