Regulation of the actin cytoskeleton in the amoeba Dictyostelium discoideum by kinases and formin-dependent activities

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Focus of this research project is the investigation (i) of regulatory principles during signal transduction to the actin cytoskeleton as mediated by kinases, and (ii) of selected formins during cytoskeleton-dependent activities. The model organism for these studies is the social amoeba Dictyostelium discoideum. (i) Ste20-like kinases are known to be involved in cytoskeletal regulation. We isolated already several knockout mutants whose aberrant phenotypes will be studied. We included in the work programme also the characterization of an unconventional kinase which might be a D. discoideum homologue of the actin-fragmin kinase from Physarum polycephalum. (ii) Our recent results on the role of the formin dDia2 for regulation of actin polymerization and formation of filopodia are the basis for further studies on additional formin activities in D. discoideum. The work programme includes the generation of single and multiple knockout mutants, the biochemical characterization of selected formin domains, and the analysis of GFP-tagged formins in wild type cells and mutants. Of special interest are the putative lipid-binding domains of D. discoideum formins ForA and dDiaS.