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Cbl interacts with multiple E2s in vitro and in cells

Mariya S Liyasova1, Ke Ma1, Donna Voeller1

  • 1Women's Malignancy Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, United States of America.

Plos One
|May 24, 2019
PubMed

Insights

Cbl proteins regulate receptor tyrosine kinases (RTKs) via ubiquitination. Multiple ubiquitin-conjugating enzymes (E2s) interact with Cbl, modulating its E3 ligase activity and influencing RTK ubiquitination in cells.

Area of Science:

  • Cellular biology
  • Molecular biology
  • Biochemistry

Background:

  • Cbl proteins are RING finger (RF) ubiquitin ligases (E3s) that negatively regulate receptor tyrosine kinases (RTKs) like EGFR and MET through ubiquitination and degradation.
  • Loss of Cbl function is linked to malignant transformation due to elevated RTK activity.

Purpose of the Study:

  • To investigate the roles of various ubiquitin-conjugating enzymes (E2s) in Cbl-mediated ubiquitination of RTKs.
  • To determine how different E2s affect Cbl E3 ligase activity and ubiquitin chain formation.

Main Methods:

  • In vitro E3 assays and yeast two-hybrid screens were employed.
  • Ubiquitination assays were performed using Cbl, E2s, and substrates like EGFR and SYK.
  • Cellular ubiquitination of EGFR was assessed.

Main Results:

  • Several E2s, including Ube2d, Ube2e, Ube2n/2v1, and Ube2w, were found to catalyze Cbl autoubiquitination.
  • Ube2d2, Ube2e1, and Ube2n/2v1 mediated Cbl-dependent ubiquitination of EGFR and SYK.
  • Phosphorylation of Cbl by Src enhanced its E3 activity.
  • Specific E2s dictated ubiquitin chain linkage types (K48, K63, or monoubiquitination).
  • Ube2d, Ube2e, and Ube2n/2v1 families contributed to EGFR ubiquitination in cells.

Conclusions:

  • Multiple E2 enzymes can interact with Cbl proteins.
  • These E2s modulate Cbl E3 ligase activity both in vitro and within cellular contexts.
  • Understanding these E2-Cbl interactions is crucial for comprehending RTK regulation and potential therapeutic strategies.

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