Objective
Recent research has identified PKD/PKCµ as a protein kinase that plays important roles in cell surface receptor and growth factor signalling as well as in stress-induced signalling by genotoxic chemotherapeutics, radiation, and oxygen radicals. Moreover, PKD/PKCµ acts as an important DAG-stimulated negative feedback regulator of plasma membrane enzymes and receptors and mediates the cross talk between different signalling systems. The recent discovery of additional kinases which belong to the PKD family and the plethora of proteins that interact with PKD/PKCµ points to a multifaceted regulation and a multifunctional role for these enzymes, with functions in processes as diverse as cell proliferation, tumor cell invasion, and regulation of Golgi vesicle fission.
The main objectives of the project are:
A. To elucidate the molecular architecture of PKD;
B.To examine how PKD activation affects p53-dependent and -independent pathways responsible for regulation of apoptosis and cell cycle checkpoints and;
C. To investigate the role of PKD in carcinogenesis.
Specific tasks of this project will be:
T1. Molecular dissection of PKD structure and function;
T2.To develop molecular tools for the detection of protein kinase D;
T3. To investigate the role of PKD in p53-upstream and p53-downstream signalling;
T4.To investigate the role of PKD in B cell signalling;
T5.To assess the role of PKD in carcinogenesis.
A wide range of biochemical, immunological, cell death, computational and molecular biology methods will be applied Among them electron and confocal microscopy, immunocytochemistry, Fluorescence-activated cell sorting, Western blot analisis, site directed mutagenesis, phosphorylation assays, phosphopeptide sequencing techniques, yeast two-hybrid system, cell cycle analysis, antisense technology. Potential results will be expected: the molecular structure of PKD of its activation will be investigated; the role of PKD in different signal cascades associated with cell growth and apoptosis, will be determined; the potential links between the activities of PKD and p53, the most frequently mutated gene in human cancer will be clarified, the role of PKD in B- and T-cell regulation will be studied; the molecular insights in PKD function will be used to design therapeutics that inhibit the action of PKD.
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Programme(s)
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Coordinator
3000 Leuven
Belgium
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