Objective Research objectives and content We will study the role of the raf-1 kinase in oocytes in the mechanisms leading to resumption of the cell cycle, causing entry into metaphase of meiosis-1, and in apoptosis. The raf-1 kinase is a central player in signal transduction in eukaryotic cells although its role in entry into metaphase of the cell cycle in oocytes is highly controversial. Current data suggests that raf-1 may have multiple roles in cell cycle progression. We intend to define precisely whether the requirement for raf-1 is solely to activate the MAP kinase cascade or whether it has an additional role in progesteronemediated meiotic maturation. These experiments will use a combination of expression in vivo of dominant-negative and constitutively active raf-1 mutants and downstream (MAP kinase cascade) protein kinases, followed by measurements of key enzyme activities in vitro. These experiments will be backed up by depletion and reconstitution experiments in extracts in vitro. Secondly we will study the mechanism of apoptosis in oocyte extracts in our working system, which responds to protein kinase signals. We will concentrate on the involvement of the raf-1 kinase, and the potential involvement of downstream peroxidation reactions, in this process. Training content (objective, benefit and expected impact) The objective is to analyse the control of fundamental cellular processes: the cell cycle and apoptosis, in a system amenable to biochemical analysis in vivo and in vitro, by combining different approaches. As a focal point the study uses a well characterized protein, the raf-1 kinase. The use of the oocyte system has been invaluable in cell cycle studies and development of an expertise by the candidate in use of this system is very important. In addition the project will give the candidate the opportunity for comparative biochemistry by testing results obtained in Xenopus oocytes in the heterologous starfish oocyte system. The potential impact of the results is very high, and would have wide-ranging implications. Links with industry / industrial relevance (22) Fields of science natural sciencesbiological sciencesbiochemistrybiomoleculesproteinsenzymes Programme(s) FP4-TMR - Specific research and technological development programme in the field of the training and mobility of researchers, 1994-1998 Topic(s) 0302 - Post-doctoral research training grants TL02 - Molecular Biology and Biochemistry Call for proposal Data not available Funding Scheme RGI - Research grants (individual fellowships) Coordinator CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE EU contribution No data Address Route de Mende 1919 34293 MONTPELLIER France See on map Total cost No data Participants (1) Sort alphabetically Sort by EU Contribution Expand all Collapse all Not available United Kingdom EU contribution No data Address See on map Total cost No data