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Related Experiment Videos

c-Ets-1 protein is hyperphosphorylated during mitosis

L F Fleischman1, A M Pilaro, K Murakami

  • 1Laboratory of Molecular Oncology, National Cancer Institute, Frederick, Maryland 21702-1201.

Oncogene
|March 1, 1993
PubMed
Summary

The study found a hyperphosphorylated form of Ets-1 protein specifically during mitosis, suggesting its role in cell cycle regulation. This modification, occurring transiently, does not affect DNA binding but is linked to cellular phosphatase activity.

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Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Ets-1 and Ets-2 are proto-oncogene products functioning as transcription factors.
  • Phosphorylation of Ets proteins is modulated by calcium-mediated signals.
  • Cell cycle regulation of transcription factors is crucial for cellular processes.

Purpose of the Study:

  • To investigate the cell cycle-dependent expression and phosphorylation of Ets-1 and Ets-2 proteins.
  • To identify the specific cell cycle phase associated with Ets-1 hyperphosphorylation.
  • To explore the functional implications of Ets-1 hyperphosphorylation on its DNA-binding activity.

Main Methods:

  • Cell synchronization using centrifugal elutriation and nocodazole-induced mitotic block.
  • Analysis of Ets-1 and Ets-2 protein isoforms via mobility shifts on gels.

Related Experiment Videos

  • Treatment with okadaic acid to assess the role of phosphatases.
  • Assessment of sequence-specific DNA-binding activity.

    • Main Results:

    • A hyperphosphorylated isoform of Ets-1 was consistently observed during mitosis across multiple cell lines.
    • This hyperphosphorylated Ets-1 isoform appeared transiently, disappearing as cells exited mitosis.
    • Okadaic acid treatment induced widespread Ets-1 hyperphosphorylation, indicating phosphatase involvement.
    • Ets-1 hyperphosphorylation did not alter its DNA-binding capabilities.
    • Enhanced phosphorylation of Ets-2 was noted, but not hyperphosphorylation.

    Conclusions:

    • Ets-1 undergoes transient hyperphosphorylation during early mitosis, potentially regulating its function.
    • Cellular phosphatase activity plays a significant role in the cell cycle-specific regulation of Ets-1.
    • The observed modifications to Ets-1 during mitosis do not impair its DNA-binding activity.