Objective
S6K is a key player in PI3K and mTOR pathways, which control cell growth, proliferation, survival and metabolism. Deregulation of S6K signalling has been found in human pathologies, such as cancer and diabetes. The family of S6Ks is represented by two forms, S6K1 and S6K2, which have cytoplasmic and nuclear splicing variants. The contribution of each S6K isoform to the regulation of gene expression (at the level of transcription and translation) and cell growth is not clear. In this proposal, the techniques of Proteome analysis and DNA microarrays will be used to study gene expression profiles in stable cell lines, which overexpress activated versions of S6K1/2. Moreover, metabolic P33-labelling of generated cells in combination with 2D electrophoresis will be used to study the profile of phosphorylated proteins. This analysis may lead to the identification by mass spectrometry of novel substrates for S6Ks or each isoform in particular. Functional studies of generated cells will complement molecular approaches described above. We are particularly interested to dissect the involvement of each isoform of S6K in the regulation of cell growth, cell survival and cytoskeletal rearrangement. Since, generated cells express high level of recombinant S6Ks fussed to EE-tag epitope, they will be used for co-purification of binding partners on matrixes containing anti-EE antibodies. To co-precipitate binding partners with endogenous S6Ks, we will employ S6K1 and S6K2 monoclonal antibodies which have been recently developed by NIS collaborators. Specifically associated proteins detected by SDS-PAGE will be identified by mass spectrometry. This study may create a strong base for further collaborative links and grant applications between participating teams, especially in testing the specificity of identified interaction(s) and their physiological importance. We believe that collaborative teams have the potential to tackle the proposed project from different angles, using modern techniques and experimental models. The proposed study has the potential to advance our understanding of S6K function in signal transduction, especially the role of its isoforms in regulating gene expression at the level of transcription and translation. Apart of fundamental importance for biological sciences, this new knowledge could be applied for development of novel diagnostic and therapeutic approaches in human pathologies, such as cancer.
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Keywords
Project’s keywords as indicated by the project coordinator. Not to be confused with the EuroSciVoc taxonomy (Fields of science)
Project’s keywords as indicated by the project coordinator. Not to be confused with the EuroSciVoc taxonomy (Fields of science)
Programme(s)
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Multi-annual funding programmes that define the EU’s priorities for research and innovation.
Topic(s)
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Calls for proposals are divided into topics. A topic defines a specific subject or area for which applicants can submit proposals. The description of a topic comprises its specific scope and the expected impact of the funded project.
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Funding Scheme
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Coordinator
LONDON
United Kingdom
The total costs incurred by this organisation to participate in the project, including direct and indirect costs. This amount is a subset of the overall project budget.