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Nuclear localization signal and phosphorylation of Serine350 specify intracellular localization of DRAK2

Hiroshi Kuwahara1, Michihiko Nishizaki, Hiroshi Kanazawa

  • 1Department of Biological Sciences, Graduate School of Science, Osaka University, Machikaneyama-cho, Toyonaka City, Osaka, Japan.

Journal of Biochemistry
|December 18, 2007
PubMed

Insights

Death-associated protein kinase-related apoptosis-inducing kinase 2 (DRAK2) shuttles between the nucleus and cytoplasm. Phosphorylation of Ser350 regulates DRAK2 nuclear import, impacting its role in apoptosis and T-cell development.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Immunology

Background:

  • DRAK2 is a serine/threonine kinase involved in apoptosis and T-cell receptor sensitivity.
  • DRAK2 exhibits dynamic subcellular localization between the nucleus and cytoplasm during cellular processes.

Purpose of the Study:

  • To investigate the mechanism regulating DRAK2's subcellular localization.
  • To identify the role of the nuclear localization signal (NLS) and its regulation in DRAK2 function.

Main Methods:

  • Site-directed mutagenesis of the putative DRAK2 NLS.
  • Confocal microscopy to track DRAK2 localization in response to stimuli (UV, PMA) and kinase expression (PKC-gamma).
  • Analysis of DRAK2 phosphorylation at Ser350.

Main Results:

  • A putative NLS sequence in DRAK2 was identified and confirmed to mediate nuclear import.
  • Phosphorylation of Ser350 by PKC-gamma inhibited DRAK2 nuclear localization.
  • UV irradiation induced nuclear accumulation of wild-type DRAK2 but not the Ser350Asp mutant.

Conclusions:

  • Phosphorylation of Ser350 is critical for regulating DRAK2 nuclear translocation.
  • This phosphorylation event likely modulates NLS activity, influencing DRAK2's role in apoptosis and T-cell development.

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