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A negative feedback loop attenuates EGF-induced morphological changes.

J B Welsh1, G N Gill, M G Rosenfeld

  • 1Department of Medicine, University of California, San Diego, La Jolla 92093.

The Journal of Cell Biology
|August 1, 1991
PubMed
Summary
This summary is machine-generated.

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Epidermal Growth Factor (EGF) receptor signaling is attenuated by protein kinase C (PKC) phosphorylation at threonine 654 (T654). This negative feedback loop regulates cell shape changes and lamellipodia retraction in response to EGF.

Area of Science:

  • Cell Biology
  • Molecular Signaling
  • Biochemistry

Background:

  • Epidermal Growth Factor (EGF) receptor tyrosine kinase activation initiates downstream signaling cascades, including protein kinase C (PKC).
  • EGF receptor signaling is subject to negative feedback mechanisms to prevent overstimulation.

Purpose of the Study:

  • To investigate the role of protein kinase C (PKC) phosphorylation of the EGF receptor in attenuating EGF receptor signaling.
  • To determine if phosphorylation of threonine 654 (T654) on the EGF receptor by PKC acts as a negative feedback mechanism.

Main Methods:

  • Utilized scanning electron microscopy (SEM) to observe EGF-induced cell morphology changes (lamellipodia retraction).
  • Expressed wild-type (WT) and mutant EGF receptors (truncated at C'973, alanine substitution at T654) in EGF receptor-deficient NR6 cells.

Related Experiment Videos

  • Investigated the effects of TPA (a PKC activator) and EGF on cell morphology and EGF receptor signaling.

    • Main Results:

    • Cells expressing WT and C'973 EGF receptors showed EGF-induced lamellipodia retraction; C'973 receptors responded faster.
    • TPA treatment blocked the EGF-induced retraction response.
    • EGF receptors with T654 mutated to alanine (A654) were resistant to TPA and exhibited more rapid, extensive EGF-induced morphologic changes.
    • Downregulation of PKC in cells with T654 EGF receptors mimicked the A654 mutation phenotype, indicating T654 phosphorylation mediates the inhibitory effect.

    Conclusions:

    • Activation of protein kinase C (PKC) and subsequent phosphorylation of the EGF receptor at threonine 654 (T654) is a key mechanism for rapid physiological attenuation of EGF receptor signaling.
    • This phosphorylation event serves as an intracellular negative feedback loop, regulating cell responses to EGF.