Drel V. Identification and characterisation of proteins which interact with SH3-containing adaptor protein Ruk in human embryonic kidney HEK293 cells

The object - protein-protein interactions, mediated by adapter protein Ruk. The aim: identification and characterization of adapter protein Rukl partners which interact with it. Methods of studies: PCR, cell culture techniques, transfection of cultured cells, immunofluorescence microscopy, affinity chromatography, Western-blotting and retardation of DNA-protein complexes in polyacrylamide gel. It was shown that the adapter protein Rukl was distributed both in cytoplasm, and in a nucleus of HEK293 and U937 cells. Was elaborated methods for affinity purification of recombinant protein Rukl Glu-tagged and endogenous protein Rukl from lysates of HEK293 cells, that based on affinity chromatography. Was found that affinity purified of recombinant and endogenous proteins Rukl associated with proteins that can interact and hydrolyze DNA. It was mapped the C-terminal “coiled-coil” region of Rukl as a domain, involved in interaction with nuclease/nucleases. The assumption concerning a biological role of proteincomplexes mediated by adapter protein Rukl with endonucleases in the control of DNA catabolism under the induction of cell apoptosis is discussed. Field of application: cell and molecular biology.