Tyrosine phosphorylation of Cbl upon epidermal growth factor (EGF) stimulation and its association with EGF receptor and downstream signaling proteins

Toru Fukazawa, Sachiko Miyake, Vimla Band, Hamid Band

Research output: Contribution to journalArticle

142 Citations (Scopus)

Abstract

We and others have shown that Cbl, the protein product of the c-cbl proto- oncogene, is an early target of tyrosine phosphorylation upon stimulation through the immune cell surface receptors, which signal through noncovalently associated cytoplasmic tyrosine kinases. Using human mammary epithelial cells that express a natural epidermal growth factor (EGF) receptor and require EGF as an essential growth factor, we demonstrate here that Cbl is a prominent target of tyrosine phosphorylation upon stimulation through the EGF receptor tyrosine kinase. Phosphorylation of Cbl was EGF dose-dependent, rapid (detectable as early as 5 s and maximal by 2 min), and relatively sustained (detectable even after 1 h). Co-immunoprecipitation studies demonstrated that Cbl became associated with the EGF receptor in an EGF-dependent manner. Cbl was basally associated with the adaptor protein growth factor receptor- binding protein 2 (Grb2), and this interaction was further enhanced by EGF stimulation; however, the interaction was entirely mediated via the Grb2 Src homology 3 (SH3) domains, suggesting that binding of Grb2 SH2 domain to EGF receptor provides one mechanism of Cbl's association with the EGF receptor. EGF stimulation also induced the association of Cbl with Src homology and collagen (Shc) protein, p85 subunit of the phosphatidylinositol 3-kinase and Crk proteins, in particular with the CrkL isoform. Interactions of Cbl with the EGF receptor and multiple downstream signaling proteins suggest a role for this proto-oncogene product in mitogenic signaling through growth factor receptor kinases.

Original languageEnglish (US)
Pages (from-to)14554-14559
Number of pages6
JournalJournal of Biological Chemistry
Volume271
Issue number24
DOIs
StatePublished - Jun 25 1996

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Phosphorylation
Epidermal Growth Factor Receptor
Epidermal Growth Factor
Tyrosine
Growth Factor Receptors
Association reactions
Carrier Proteins
Proteins
src Homology Domains
Protein-Tyrosine Kinases
Proto-Oncogene Proteins c-cbl
Phosphatidylinositol 3-Kinase
Proto-Oncogenes
Oncogene Proteins
Protein Subunits
Cell Surface Receptors
Immunoprecipitation
Intercellular Signaling Peptides and Proteins
Protein Isoforms
Breast

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Tyrosine phosphorylation of Cbl upon epidermal growth factor (EGF) stimulation and its association with EGF receptor and downstream signaling proteins. / Fukazawa, Toru; Miyake, Sachiko; Band, Vimla; Band, Hamid.

In: Journal of Biological Chemistry, Vol. 271, No. 24, 25.06.1996, p. 14554-14559.

Research output: Contribution to journalArticle

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