Research objectives and content
The objective of the research is to investigate the role of the recently discovered Erk- and p38 kinase-activated kinases Mnk1 and Mnk2 in the regulation of the physiology of eukaryotic cells. Both Mnk1 and Mnk2 are candidates for key mediators of the actions of multiple signaling pathways. The main goals of the research are:
1. Identifying substrates of Mnk1/2 by: I. screening 1 phage DNA expression libraries with purified active Mnk1/2 in a solid-phase phosphorylation assay. II. the yeast two-hybrid method. 2. Determination under which conditions Mnk1/2 are activated Various cell type/stimulus combinations will be used to study the activation of Mnk1 and Mnk2. For example, we will use human embryonic kidney 293 cells, as data on the activation of Mnk1 are already available and they are readily transfected.
3. Determination of cellular roles of Mnk1/2.By transfection of cells with dominant negative or constitutively active mutants of cells we will study the effect of the kinases on the activity or phosphorylation of substrates, and on cell growth etc..
4. Determination of the role of Mnk2 in eIF4E phosphorylation.Several approaches will be employed to establish whether Mnk2 is, like Mnk1, able to phosphoryate the key translation initiation factoreIF4E.
Training content (objective, benefit and expected impact)
The proposed research will substantially broaden the applicant's expertisee. The applicant will learn a range of new techniques, such as protein purification/characterisation and use of dominant negative and constitutively active mutants. This research will especially increase his knowledge and experience in the fast-growing field of signal transduction. The benefits of the proposed research are mainly of fundamental scientific value. The results may help in understanding mechanisms underlying disease states (e.g. cancer) or growth defects.
Links with industry / industrial relevance (22)
The group has several links with Pfizer Central Research in exploring the roles of intracellular signalling pathways in regulating cell function including protein synthesis and cell growth. These include the p38 MAP kinase pathway which regulates Mnk1, the subject of this study.