Cbl-mediated negative regulation of platelet-derived growth factor receptor-dependent cell proliferation. A critical role for Cbl tyrosine kinase-binding domain

Sachiko Miyake, Karen P. Mullane-Robinson, Nancy L. Lill, Patrice Douillard, Hamid Band

Research output: Contribution to journalArticle

155 Citations (Scopus)

Abstract

The Cbl proto-oncogene product has emerged as a novel negative regulator of receptor and non-receptor tyrosine kinases. Our previous observations that Cbl overexpression in NIH3T3 cells enhanced the ubiquitination and degradation of the platelet-derived growth factor receptor-α (PDGFRα) and that the expression of oncogenic Cbl mutants up-regulated the PDGFRα signaling machinery strongly suggested that Cbl negatively regulates PDGFRα signaling. Here, we show that, similar to PDGFRα, selective stimulation of PDGFRβ induces Cbl phosphorylation, and its physical association with the receptor. Overexpression of wild type Cbl in NIH3T3 cells led to an enhancement of the ligand-dependent ubiquitination and subsequent degradation of the PDGFRβ, as observed with PDGFRα. We show that Cbl-dependent negative regulation of PDGFRα and results in a reduction of PDGF-induced cell proliferation and protection against apoptosis. A point mutation (G306E) that inactivates the tyrosine kinase binding domain in the N-terminal transforming region of Cbl compromised the PDGF-inducible tyrosine phosphorylation of Cbl although this mutant could still associate with the PDGFR. More importantly, the G306E mutation abrogated the ability of Cbl to enhance the ligand- induced ubiquitination and degradation of the PDGFR and to inhibit the PDGF- dependent cell proliferation and protection from apoptosis. These results demonstrate that Cbl can negatively regulate PDGFR-dependent biological responses and that this function requires the conserved tyrosine kinase binding domain of Cbl.

Original languageEnglish (US)
Pages (from-to)16619-16628
Number of pages10
JournalJournal of Biological Chemistry
Volume274
Issue number23
DOIs
StatePublished - Jun 4 1999

Fingerprint

Platelet-Derived Growth Factor Receptors
Cell proliferation
Protein-Tyrosine Kinases
Cell Proliferation
Ubiquitination
Phosphorylation
Cytoprotection
Degradation
Apoptosis
Ligands
Proto-Oncogenes
Oncogene Proteins
Point Mutation
Machinery
Tyrosine
Association reactions

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Cbl-mediated negative regulation of platelet-derived growth factor receptor-dependent cell proliferation. A critical role for Cbl tyrosine kinase-binding domain. / Miyake, Sachiko; Mullane-Robinson, Karen P.; Lill, Nancy L.; Douillard, Patrice; Band, Hamid.

In: Journal of Biological Chemistry, Vol. 274, No. 23, 04.06.1999, p. 16619-16628.

Research output: Contribution to journalArticle

@article{04b7a0f7a27f453ba706a8baed283976,
title = "Cbl-mediated negative regulation of platelet-derived growth factor receptor-dependent cell proliferation. A critical role for Cbl tyrosine kinase-binding domain",
abstract = "The Cbl proto-oncogene product has emerged as a novel negative regulator of receptor and non-receptor tyrosine kinases. Our previous observations that Cbl overexpression in NIH3T3 cells enhanced the ubiquitination and degradation of the platelet-derived growth factor receptor-α (PDGFRα) and that the expression of oncogenic Cbl mutants up-regulated the PDGFRα signaling machinery strongly suggested that Cbl negatively regulates PDGFRα signaling. Here, we show that, similar to PDGFRα, selective stimulation of PDGFRβ induces Cbl phosphorylation, and its physical association with the receptor. Overexpression of wild type Cbl in NIH3T3 cells led to an enhancement of the ligand-dependent ubiquitination and subsequent degradation of the PDGFRβ, as observed with PDGFRα. We show that Cbl-dependent negative regulation of PDGFRα and results in a reduction of PDGF-induced cell proliferation and protection against apoptosis. A point mutation (G306E) that inactivates the tyrosine kinase binding domain in the N-terminal transforming region of Cbl compromised the PDGF-inducible tyrosine phosphorylation of Cbl although this mutant could still associate with the PDGFR. More importantly, the G306E mutation abrogated the ability of Cbl to enhance the ligand- induced ubiquitination and degradation of the PDGFR and to inhibit the PDGF- dependent cell proliferation and protection from apoptosis. These results demonstrate that Cbl can negatively regulate PDGFR-dependent biological responses and that this function requires the conserved tyrosine kinase binding domain of Cbl.",
author = "Sachiko Miyake and Mullane-Robinson, {Karen P.} and Lill, {Nancy L.} and Patrice Douillard and Hamid Band",
year = "1999",
month = "6",
day = "4",
doi = "10.1074/jbc.274.23.16619",
language = "English (US)",
volume = "274",
pages = "16619--16628",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "23",

}

TY - JOUR

T1 - Cbl-mediated negative regulation of platelet-derived growth factor receptor-dependent cell proliferation. A critical role for Cbl tyrosine kinase-binding domain

AU - Miyake, Sachiko

AU - Mullane-Robinson, Karen P.

AU - Lill, Nancy L.

AU - Douillard, Patrice

AU - Band, Hamid

PY - 1999/6/4

Y1 - 1999/6/4

N2 - The Cbl proto-oncogene product has emerged as a novel negative regulator of receptor and non-receptor tyrosine kinases. Our previous observations that Cbl overexpression in NIH3T3 cells enhanced the ubiquitination and degradation of the platelet-derived growth factor receptor-α (PDGFRα) and that the expression of oncogenic Cbl mutants up-regulated the PDGFRα signaling machinery strongly suggested that Cbl negatively regulates PDGFRα signaling. Here, we show that, similar to PDGFRα, selective stimulation of PDGFRβ induces Cbl phosphorylation, and its physical association with the receptor. Overexpression of wild type Cbl in NIH3T3 cells led to an enhancement of the ligand-dependent ubiquitination and subsequent degradation of the PDGFRβ, as observed with PDGFRα. We show that Cbl-dependent negative regulation of PDGFRα and results in a reduction of PDGF-induced cell proliferation and protection against apoptosis. A point mutation (G306E) that inactivates the tyrosine kinase binding domain in the N-terminal transforming region of Cbl compromised the PDGF-inducible tyrosine phosphorylation of Cbl although this mutant could still associate with the PDGFR. More importantly, the G306E mutation abrogated the ability of Cbl to enhance the ligand- induced ubiquitination and degradation of the PDGFR and to inhibit the PDGF- dependent cell proliferation and protection from apoptosis. These results demonstrate that Cbl can negatively regulate PDGFR-dependent biological responses and that this function requires the conserved tyrosine kinase binding domain of Cbl.

AB - The Cbl proto-oncogene product has emerged as a novel negative regulator of receptor and non-receptor tyrosine kinases. Our previous observations that Cbl overexpression in NIH3T3 cells enhanced the ubiquitination and degradation of the platelet-derived growth factor receptor-α (PDGFRα) and that the expression of oncogenic Cbl mutants up-regulated the PDGFRα signaling machinery strongly suggested that Cbl negatively regulates PDGFRα signaling. Here, we show that, similar to PDGFRα, selective stimulation of PDGFRβ induces Cbl phosphorylation, and its physical association with the receptor. Overexpression of wild type Cbl in NIH3T3 cells led to an enhancement of the ligand-dependent ubiquitination and subsequent degradation of the PDGFRβ, as observed with PDGFRα. We show that Cbl-dependent negative regulation of PDGFRα and results in a reduction of PDGF-induced cell proliferation and protection against apoptosis. A point mutation (G306E) that inactivates the tyrosine kinase binding domain in the N-terminal transforming region of Cbl compromised the PDGF-inducible tyrosine phosphorylation of Cbl although this mutant could still associate with the PDGFR. More importantly, the G306E mutation abrogated the ability of Cbl to enhance the ligand- induced ubiquitination and degradation of the PDGFR and to inhibit the PDGF- dependent cell proliferation and protection from apoptosis. These results demonstrate that Cbl can negatively regulate PDGFR-dependent biological responses and that this function requires the conserved tyrosine kinase binding domain of Cbl.

UR - http://www.scopus.com/inward/record.url?scp=0033523010&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033523010&partnerID=8YFLogxK

U2 - 10.1074/jbc.274.23.16619

DO - 10.1074/jbc.274.23.16619

M3 - Article

C2 - 10347229

AN - SCOPUS:0033523010

VL - 274

SP - 16619

EP - 16628

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 23

ER -