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c-Cbl ubiquitinates the EGF receptor at the plasma membrane and remains receptor associated throughout the endocytic

A A de Melker1, G van der Horst, J Calafat

  • 1Division of Cellular Biochemistry, The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands.

Journal of Cell Science
|August 9, 2001
PubMed

Insights

The Cbl protein (c-Cbl) binds and ubiquitinates the epidermal growth factor (EGF) receptor at the plasma membrane, facilitating its entry into clathrin-coated pits and subsequent intracellular sorting.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Cbl family proteins are key negative regulators of receptor tyrosine kinase (RTK) signaling.
  • Their function relies on a Ring finger domain, enabling interaction with ubiquitin conjugating enzymes.
  • Cbl proteins act as E3 ubiquitin ligases, targeting RTKs for downregulation.

Purpose of the Study:

  • To investigate the precise stage in the endocytic pathway where Cbl proteins interact with RTKs.
  • To elucidate the role of c-Cbl in the internalization and trafficking of the epidermal growth factor (EGF) receptor.

Main Methods:

  • Utilized low-temperature and dynamin mutant approaches to study receptor dynamics.
  • Employed confocal microscopy and immunogold electron microscopy for high-resolution imaging.
  • Investigated the association and ubiquitination of the EGF receptor by c-Cbl.

Main Results:

  • Demonstrated that c-Cbl associates with and ubiquitinates the activated EGF receptor at the plasma membrane before endocytosis.
  • Showed c-Cbl association with the EGF receptor persists throughout clathrin-mediated endocytosis.
  • Observed colocalization of c-Cbl and the EGF receptor in internal vesicles of multivesicular endosomes.

Conclusions:

  • c-Cbl plays a crucial role in the clathrin-mediated endocytosis of tyrosine kinase receptors.
  • c-Cbl mediates the ubiquitination of the EGF receptor at the plasma membrane, initiating its downregulation.
  • c-Cbl is involved in both the internalization and intracellular sorting of RTKs.

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