Identification of the interacting partners of CD3 delta

Official URL: http://risc01.sabanciuniv.edu/record=b1427168 (Table of Contents)

In this study, we identify the CD3δ interacting proteins with a special yeast two hybrid screen which uses the split-ubiquitin system. CD3δ is a subunit of the T cell receptor, which is important not only for immune function but also for T lymphocyte development. CD3δ is necessary for T cell development, therefore identifying its role in TCR signaling and function is important for understanding the immune system. In order to find out the proteins that bind to both the cytoplasmic and transmembrane domains of CD3δ, we used a membrane based- split ubiquitin yeast two hybrid system. We over expressed the putative interactors in mammalian tissue culture cells to confirm validity and specificity of the interaction. Although we could detect the interaction by yeast two hybrid screening, we could not confirm the same interactions in mammalian cells. We also attempted to express different T cell receptor subunits individually in yeast cells, and attempted to demonstrate the interaction of these protein subunits in our yeast two hybrid interaction system. Using this technique we show that CD3δε heterodimers can assemble in yeast cells.