Biopolym. Cell. 2002; 18(4):312-318.
Cell Biology
Subcellular localization of adapter protein Ruk, in HEK293 cells
1Drel V. R., 1Palyvoda O. Yu., 1Shuvajeva G. Yu., 1Ihumenceva N. I., 1Kit Yu. Ya., 2, 3Gout I. T., 4Buchman V. L., 1Drobot L. B.
  1. Institute of Cell Biology, NAS of Ukraine
    14/16, Drahomanov Str., Lviv, Ukraine, 79005
  2. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680
  3. Ludwig Institute for Cancer Research
    91 Riding house str., London, UK, WIP8BT
  4. The Universuty of Edinburgh
    Edinburgh, UK

Abstract

Subcellular localization of adapter protein Ruk1 has been investigated in human embryonic kidney HEK293 cells transfected with pRc/CMV2/Ruk1-Glu-tag. Using immunofluorescence microscopy, it was shown that the recombinant protein was distributed diffusely in both cytoplasm and nucleus, but with punctated structures in the nucleus, which correspond in all probability to the nucleolus. The Ruk1 localization in the nucleus was confirmed by Western blotting of nucleic extracts prepared from transfected HEK293 cells using monoclonal anti-Glu-tag antibodies, as well as from nontransfected HEK293 and U937 cells using polyclonal anti-Ruk antibodies. The ability of affine purified Ruk1 Glu-tagged preparation to bind to DNA from calf thymus, but not to Escherichia coli DNA, was revealed by the immunodot-blot analysis. The nuclear localization of Ruk1 suggests that this adapter protein is involved in some new, yet unrecognized functions, in the eukaryotic cell nucleus.

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