Functional analysis of a calcium sensor protein/serine-threonine kinase signalling network

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Calcium signals regulate a multitude of biological responses and cellular processes in eukaryotic organisms. Although considerable progress has been made in elucidating physiological processes involving calcium signals in plants, the identity of most of the components involved and the mechanisms generating specificity in this signalling cascades are only beginning to be understood. This project aims to comprehensively explore the mechanisms of a novel calcium signalling system from Arabidopsis. Based on the recent identification of a specific group of serine-threonine protein kinases (AtCIPKs) as interacting targets of the Arabidopsis calcineurin B-like calcium sensor proteins (AtCBLs) this project is focused on the functional characterisation of the CBL-CIPK protein-protein interaction network and its contribution to integrate and specifically decode the various calcium signals in plant cells. The proposed research addresses the folowing main objectives: (1) A thorough molecular characterisation of the CBL and CIPK protein families by their expression, subcelluar localisation, biochemical properties and interaction specificy. (2) The identification of the specific physiological processes by the diffrential calcium sensor/kinase complex formation. This approach will rely on the isolation and detailed phenotypical physiological and genetic charakterisation of mutant lines generated by reserve genetic approaches as well as on interaction analyses by yeast two-hybrid assays. (3) The identification and analyses of proteins regulated by the CBL-CIPK complexes by means of large scale two- and three-hybrid screens.