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Analysis of nucleo-cytoplasmic shuttling of the proto-oncogene SET/I2PP2A

B Daniel Lam1, Eloise C Anthony, Peter L Hordijk

  • 1Department of Molecular Cell Biology, Sanquin Research and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands.

Cytometry. Part a : the Journal of the International Society for Analytical Cytology
|October 13, 2011
PubMed

Insights

Researchers developed a new method to track the SET protein

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Oncology

Background:

  • SET/I2PP2A is a nuclear proto-oncogene implicated in acute myeloid leukemia.
  • SET inhibits the phosphatase PP2A and may regulate cell migration.
  • Rac1 GTPase was previously proposed to recruit SET from the nucleus to the plasma membrane.

Purpose of the Study:

  • To develop and validate a method for analyzing nucleo-cytoplasmic shuttling of the SET protein in live cells.
  • To investigate the dynamics of SET protein localization under various cellular conditions.
  • To assess the impact of SET protein phosphorylation on its nuclear export.

Main Methods:

  • Generation of fluorescent SET protein variants for live-cell imaging.
  • Utilizing 4D confocal microscopy to capture dynamic protein localization.
  • Development of an image analysis technique to quantify nuclear exit frequency.
  • Analysis of wild-type SET and phosphomimetic/mutant variants (S9E, S9A).

Main Results:

  • SET protein exhibits nucleo-cytoplasmic shuttling, with a fraction translocating to the cytosol.
  • Nuclear exit of SET was observed in spreading and dividing cells.
  • The frequency of nuclear exit was enhanced in the Ser-9 phosphomimetic mutant (S9E).
  • The developed image analysis method successfully quantified SET nuclear export dynamics.

Conclusions:

  • A novel method for analyzing SET protein nucleo-cytoplasmic shuttling in live cells has been established.
  • SET protein dynamics are influenced by phosphorylation at Ser-9.
  • This method is applicable for studying the dynamic localization of other signaling proteins.
  • Understanding SET shuttling may provide insights into its role in cancer and cell migration.

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