An essential role of ubiquitination in Cbl-mediated negative regulation of the Src-family kinase Fyn

Navin Rao, Amiya K. Ghosh, Pengcheng Zhou, Patrice Douillard, Christopher E. Andoniou, Hamid Band

Research output: Contribution to journalArticle

11 Scopus citations

Abstract

The Cbl family of ubiquitin ligases function as negative regulators of activated receptor tyrosine kinases by facilitating their ubiquitination and subsequent lysosomal targeting. Here, we have investigated the role of Cbl ubiquitin ligase activity in the negative regulation of a non-receptor tyrosine kinase, the Src-family kinase Fyn. Using mouse embryonic fibroblasts from Cbl+/+ and Cbl-/- mice, we demonstrate that endogenous Cbl mediates the ubiquitination of Fyn and dictates the rate of Fyn turnover. By analyzing CHO-TS20 cells with a temperature-sensitive ubiquitin activating enzyme, we demonstrate that intact cellular ubiquitin machinery is required for Cbl-induced degradation of Fyn. Analyses of Cbl mutants, with mutations in or near the RING finger domain, in 293T cells revealed that the ubiquitin ligase activity of Cbl is essential for Cbl-induced degradation of Fyn by the proteasome pathway. Finally, use of a SRE-luciferase reporter demonstrated that Cbl-dependent negative regulation of Fyn function requires the region of Cbl that mediates the ubiquitin ligase activity. Given the conservation of structure between various Src-family kinases and the ability of Cbl to interact with multiple members of this family, Cbl-dependent ubiquitination could serve a general role to negatively regulate activated Src-family kinases.

Original languageEnglish (US)
Pages (from-to)29-39
Number of pages11
JournalSignal Transduction
Volume2
Issue number1-2
DOIs
StatePublished - 2002

Keywords

  • Degradation
  • Regulation
  • Tyrosine kinase
  • Ubiquitin

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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