A complex of Shc and Ran-GTPase localises to the cell nucleus

  • R. George
  • H.-L. Chan
  • Z. Ahmed
  • K. M. Suen
  • C. N. Stevens
  • J. A. Levitt
  • K. Suhling
  • J. Timms
  • J. E. Ladbury
Research Article

Abstract.

The three isoforms of the adaptor protein Shc play diverse roles in cell signalling. For example, the observation of p46 Shc in the nuclei of hepatocellular carcinoma cells suggests a function quite distinct from the better characterised cytoplasmic role. Ligands responsible for the transport of various Shc isoforms into organelles such as the nucleus have yet to be reported. To identify such ligands a far western approach was used to determine the p52 Shc interactome. The Ran-GTPase nuclear transport protein was identified and found to bind to p52 Shc in vitro with low micromolar affinity. Co-immunoprecipitation, pull down and fluorescence lifetime imaging microscopy experiments in stable cells confirmed cellular interaction and nuclear localisation. The nuclear transport factor protein NTF2, which functions in cohort with Ran, was shown to form a complex with both RAN and Shc, suggesting a mechanism for Shc entry into the nucleus as part of a tertiary complex.

Keywords.

Nuclear localisation protein complex FLIM nuclear transport proteomics 

Supplementary material

18_2009_8667_MOESM1_ESM.pdf (76 kb)
(PDF 77 kb)

Copyright information

© Birkhäuser Verlag, Basel 2009

Authors and Affiliations

  • R. George
    • 1
  • H.-L. Chan
    • 2
  • Z. Ahmed
    • 1
  • K. M. Suen
    • 1
  • C. N. Stevens
    • 1
  • J. A. Levitt
    • 3
  • K. Suhling
    • 3
  • J. Timms
    • 2
  • J. E. Ladbury
    • 1
  1. 1.Department of Biochemistry and Molecular BiologyUniversity College LondonLondonUK
  2. 2.Cancer Proteomics Laboratory, EGA Institute for Women’s HealthUniversity College LondonLondonUK
  3. 3.Department of PhysicsKings College LondonLondonUK

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