Gamma-tubulin is involved in the nucleation of microtubules from microtubule organizing centers. Recent data document that gamma-tubulin is also associated with membranes and interacts with Src family protein tyrosine kinase Fyn and p85? regulatory subunit of phosphoinositide 3-kinase (PI3K), that are involved in signal transduction. The aim of this research project is to elucidate the role of Fyn and PI3K in regulation of gamma-tubulin functions in mast cell signaling. To this end we will characterize protein complexes containing gamma-tubulin and signaling molecules, prepare p85? functional domains and their parts for pull down assays, and analyze gamma-tubulin forms in activated cells. Immunofluorescence microscopy and live cell imaging will be used for evaluation of microtubule nucleation and microtubule dynamics in cells. Both wild-type cells and cells with kinases silenced by RNAi will be used. These studies could lead to a better understanding of gamma-tubulin functions in normal cells and under various disease conditions. (en)
Cílem tohoto projektu je pomocí molekulárně biologických přístupů a mikroskopických technik porozumět úloze klíčových signálních molekul při regulaci nukleace mikrotubulů v průběhu aktivace žírných buněk. Studium povede k hlubšímu pochopení tvorby mikrotubulů za normálních a patologických stavů.