Breus O. Functional links of CoA Synthase with components of cell signal transduction pathways

The object - molecular mechanisms of signal transduction system functioning and their interlinks with metabolic processes. The aim of the work was identification of functional links between CoA Synthase and components of cellular signal transduction pathways. The methods applied - bioinformatics, affinity chromatography, transient and stable cell transformation with cDNA constructs and siRNA, proteins immunoprecipitation and Western-blot analysis, subcellular fractionation, confocal immunofluorescent microscopy, colonies-forming test in semisolid agar etc. The results and novelity - new isoform of CoA Synthase have been identified. Signalling proteins which interacts with CoA Synthase in mammalian cells have been identified - protein tyrosine phosphatase Shp2 and regulatory subunit of PI3K, p85. It is demonstrated for the first time, that a fraction of CoA Synthase is undergone to tyrosine phosphorylation in mammalian cells and such phosphorylation may influens its PPAT activity. Have been demonstrated that expression level of CoA Synthase influences activity of PI3K-dependent pathway and tyrosine kinases. It is demonstrated for the first time that expression level of CoA Synthase influences phenotypic manifestations of cellular malignant transformation such as an ancorage independent growth and survival in serum depleted conditions in in vitro culture.