Expression of I2PP2A, an inhibitor of protein phosphatase 2A, induces c-Jun and AP-1 activity

S W Al-Murrani1, J R Woodgett, Z Damuni

  • 1Department of Cellular and Molecular Physiology H166, The Pennsylvania State University College of Medicine, Milton S. Hershey Medical Centre, P.O. Box 850, Hershey, PA 17033, USA.

Insights

Transient expression of I2PP2A, a protein phosphatase 2A inhibitor, increased c-Jun levels and DNA binding. Protein phosphatase 2A catalytic subunit (PP2AC) decreased c-Jun, suggesting PP2A regulates c-Jun phosphorylation.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Protein phosphatase 2A (PP2A) is a key regulator of cellular processes.
  • The proto-oncogene c-Jun is involved in cell proliferation and differentiation.
  • PP2A's role in regulating c-Jun activity and phosphorylation is not fully understood.

Purpose of the Study:

  • To investigate the effect of I2PP2A and PP2AC on c-Jun concentration, DNA binding, and transcriptional activity.
  • To explore the role of c-Jun phosphorylation at Ser63 and Ser73 in mediating these effects.

Main Methods:

  • Transient expression of I2PP2A and PP2AC in HEK-293 cells.
  • Western blotting to assess c-Jun concentration and phosphorylation status (phospho-Ser63 and Ser73).
  • Densitometric analysis to quantify protein levels and phosphorylation.

Main Results:

  • I2PP2A expression increased c-Jun concentration and DNA binding.
  • PP2AC expression decreased c-Jun concentration and DNA binding.
  • I2PP2A enhanced activator protein-1 transcriptional activity, an effect reduced by PP2AC.
  • Changes in c-Jun phosphorylation at Ser63 correlated with these effects.

Conclusions:

  • I2PP2A acts as a functional inhibitor of PP2A in intact cells.
  • PP2A regulates c-Jun concentration, DNA binding, and transcriptional activity.
  • PP2A may exhibit site selectivity in phosphorylating c-Jun.

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