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Related Experiment Videos

Sp1 phosphorylation by Erk 2 stimulates DNA binding

J L Merchant1, M Du, A Todisco

  • 1Department of Internal Medicine, University of Michigan, Ann Arbor, Michigan, 48109-0650, USA.

Biochemical and Biophysical Research Communications
|January 27, 1999
PubMed
Summary

Epidermal Growth Factor (EGF) signaling activates gastrin gene expression by targeting the Sp1 transcription factor. This process involves the ras-Erk pathway, where Sp1 phosphorylation regulates its binding to the gastrin gene promoter.

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Area of Science:

  • Molecular Biology
  • Cell Signaling
  • Gene Regulation

Background:

  • Epidermal Growth Factor (EGF) activates gene expression via pathways like ras-Erk.
  • EGF receptor activation stimulates gastrin gene expression through a GC-rich element (gERE) that binds Sp1.
  • The ras-Erk pathway's potential to target the gERE and Sp1 was previously unexplored.

Purpose of the Study:

  • To investigate if the ras-Erk signal transduction cascade targets the gERE element.
  • To determine if Erk 2 can phosphorylate Sp1, influencing its binding to the gERE.
  • To elucidate the role of Sp1 phosphorylation in EGF-inducible gastrin gene expression.

Main Methods:

  • Cotransfection experiments were used to assess ras and Erk 2 activation on the gERE.
  • The Mek 1 kinase inhibitor PD98059 was employed to block EGF-inducible promoter activity.

Related Experiment Videos

  • In vitro assays examined the effect of Erk 2 phosphorylation and dephosphorylation on Sp1 binding.

    • Main Results:

    • Ras and Erk 2 activation were shown to target the gERE element.
    • PD98059 inhibited approximately 50% of EGF-inducible gastrin promoter activity.
    • Erk 2 phosphorylation enhanced Sp1 binding to the gERE, while dephosphorylation reduced it.

    Conclusions:

    • Inducible Sp1 binding is regulated by its phosphorylation status.
    • The ras-Erk signaling cascade partially mediates gastrin promoter activation by targeting Sp1.
    • This study reveals a novel mechanism linking EGF signaling to Sp1-mediated gene regulation.