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PMID: 11726515 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Tyrosine phosphorylation of Grb2 by Bcr/Abl and epidermal growth factor receptor: a novel regulatory mechanism for tyrosine kinase signaling.

The EMBO journal ·Vol. 20 ·No. 23 ·2001-12-03 ·Pages 6793-804

Li S, Couvillon AD, Brasher BB, Van Etten RA

Abstract

Growth factor receptor-binding protein-2 (Grb2) plays a key role in signal transduction initiated by Bcr/Abl oncoproteins and growth factors, functioning as an adaptor protein through its Src homology 2 and 3 (SH2 and SH3) domains. We found that Grb2 was tyrosine-phosphorylated in cells expressing BCR/ABL and in A431 cells stimulated with epidermal growth factor (EGF). Phosphorylation of Grb2 by Bcr/Abl or EGF receptor reduced its SH3-dependent binding to Sos in vivo, but not its SH2-dependent binding to Bcr/Abl. Tyr209 within the C-terminal SH3 domain of Grb2 was identified as one of the tyrosine phosphorylation sites, and phosphorylation of Tyr209 abolished the binding of the SH3 domain to a proline-rich Sos peptide in vitro. In vivo expression of a Grb2 mutant where Tyr209 was changed to phenylalanine enhanced BCR/ABL-induced ERK activation and fibroblast transformation, and potentiated and prolonged Grb2-mediated activation of Ras, mitogen-activated protein kinase and c-Jun N-terminal kinase in response to EGF stimulation. These results suggest that tyrosine phosphorylation of Grb2 is a novel mechanism of down-regulation of tyrosine kinase signaling.

MeSH Terms
3T3 Cells Adaptor Proteins, Signal Transducing Animals Binding Sites Blotting, Western Cell Line Cell Line, Transformed DNA Mutational Analysis Dose-Response Relationship, Drug Down-Regulation Enzyme Activation ErbB Receptors/metabolism Fibroblasts/metabolism Fusion Proteins, bcr-abl/metabolism GRB2 Adaptor Protein Humans JNK Mitogen-Activated Protein Kinases K562 Cells MAP Kinase Signaling System Mass Spectrometry Mice Mitogen-Activated Protein Kinases/metabolism Mutation Peptide Mapping Phosphopeptides/chemistry Phosphorylation Precipitin Tests Protein Binding Protein Structure, Tertiary Proteins/metabolism Signal Transduction Son of Sevenless Protein, Drosophila/metabolism Time Factors Transfection Transformation, Genetic Tumor Cells, Cultured Tyrosine/metabolism ras Proteins/metabolism src Homology Domains
Chemicals
Adaptor Proteins, Signal Transducing GRB2 Adaptor Protein GRB2 protein, human Grb2 protein, mouse Phosphopeptides Proteins Son of Sevenless Protein, Drosophila Tyrosine ErbB Receptors Fusion Proteins, bcr-abl JNK Mitogen-Activated Protein Kinases Mitogen-Activated Protein Kinases ras Proteins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Li S
Center for Blood Research, Department of Genetics, Harvard Medical School, Boston, MA 02115, USA.
Couvillon A D
Brasher B B
Van Etten R A
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/20.23.6793
Published
2001-12-03
Pages
6793-804
Language
English
Region
England
NLM ID
8208664
PMCID
PMC125747
Subset
IM
Grants
NCI NIH HHS · R01 CA090576 · United States
NCI NIH HHS · CA72465 · United States
NCI NIH HHS · CA90576 · United States
Databases
PDB
4G10
External Links
PubMed source DOI: 10.1093/emboj/20.23.6793 PMC Full Text View journal details
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