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Content archived on 2024-04-30

The role of the raf-1 kinase in cell cycle control and apoptosis in oocytes


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)

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Route de Mende 1919

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