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Information × Registration Number 0207U005289, 0104U000724 , R & D reports Title Role of SH3-containing adaptor proteins Ruk/CIN85/SETA in the control of signalling networks in normal and transformed cells popup.stage_title Head Drobot Lyudmyla Borysivna, Доктор біологічних наук Registration Date 23-03-2007 Organization Institute of cell biology, National Academy of Sciences of Ukraine popup.description2 3. The objeсt: protein-protein interactions mediated by the adaptor protein Ruk/CIN85/SETA. The role of Ruk/CIN85/SETA isoforms in regulation of cell apoptosis. The aim: to elucidate intra- and extramolecular mechanisms of interactions mediated by the Ruk domains and motifs. To study changes of expression level and subcellular localization of Ruk/CIN85/SETA in cell apoptotic answer. To generate stable lines of human embrionic kidney HEK293 and human breast adenocarcinoma MCF7 with overexpression of full-lenth form of Ruk/CIN85 adaptor protein and to study its biological consequences. Methods: expression of recombinant proteins in bacterial and mammalian systems, purification of proteins using affinity chromatography, cell culture methods, electrophoresis of PAAG, DNA-, RNA-electrophoreses, Western blot analyses, immunoprecipitation, immunofluorescense microscopy.Theoretical and practical results: The study demonstrates that SH3A and less SH3B Ruk domains cooperatively interact with own proline-rich motifs while coil-coiled domain provides to isoform oligomerization. SH3 domain doesn't take part in intramolecular interactions, but can plays the regulatory role on general Ruk conformation. The features of Ruk/CIN85 expression are found out in cell lines with different sensitivity to influence of apoptotic inductors: A549 (human lung carcinoma), MCF7wt (human breast adenocarcinoma), L1210wt (mouse lymphocytic leukemia). It was shown that adaptor protein Ruk translocates to the perinuclear zone in cell line C6 (rat glioma) after apoptosis induction by cyclosporine A. As a result of experimental works the stable cell lines HEK293 and MCF7were obtained with different overexpression of full-lenth form of Ruk/CIN85 adaptor protein and were studied its biological consequences. Product Description popup.authors popup.nrat_date 2020-04-02 Close