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PMID: 8262059 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Mutation of Tyr697, a GRB2-binding site, and Tyr721, a PI 3-kinase binding site, abrogates signal transduction by the murine CSF-1 receptor expressed in Rat-2 fibroblasts.

The EMBO journal ·Vol. 12 ·No. 13 ·1993-12-15 ·Pages 5161-72

van der Geer P, Hunter T

Abstract

The receptor for the myeloid cell growth factor colony stimulating factor 1 (CSF-1) is a protein tyrosine kinase that is closely related to the PDGF receptor. Ligand binding results in kinase activation and autophosphorylation. Three autophosphorylation sites, Tyr697, Tyr706 and Tyr721, have been mapped to the kinase insert domain. Deletion of the entire kinase insert domain completely abrogates signal transduction by the CSF-1 receptor expressed in Rat-2 fibroblasts. To investigate the function of individual phosphorylation sites present in the CSF-1 receptor kinase insert domain, a number of phosphorylation site mutants were expressed in Rat-2 fibroblasts. Mutation of either Tyr697 or Tyr721 compromised signal transduction by the CSF-1 receptor. A mutant receptor, in which both Tyr697 and Tyr721 were replaced by phenylalanine, has lost all ability to induce changes in morphology or to increase cell growth rate in response to CSF-1. Tyr721 has been identified recently as the binding site for PI 3-kinase. Here we report that GRB2 associates with the CSF-1 receptor upon ligand binding. The phosphorylation on tyrosine of SHC and several other GRB2-associated proteins increased upon stimulation with CSF-1. Tyr697 was identified as a binding site for GRB2. We suggest that PI 3-kinase, GRB2 and some of the GRB2-associated proteins could play an important role in signal transduction by the CSF-1 receptor.

MeSH Terms
Adaptor Proteins, Signal Transducing Amino Acid Sequence Animals Binding Sites Cell Size GRB2 Adaptor Protein Ligands Macrophage Colony-Stimulating Factor/pharmacology Mice Molecular Sequence Data Mutagenesis, Site-Directed Peptides/chemistry Phosphatidylinositol 3-Kinases Phosphorylation Phosphotransferases (Alcohol Group Acceptor)/metabolism Protein Binding Proteins/metabolism Rats Receptor Protein-Tyrosine Kinases/metabolism Receptor, Macrophage Colony-Stimulating Factor/chemistry Recombinant Proteins Signal Transduction Structure-Activity Relationship Tyrosine/chemistry
Chemicals
Adaptor Proteins, Signal Transducing GRB2 Adaptor Protein Grb2 protein, mouse Grb2 protein, rat Ligands Peptides Proteins Recombinant Proteins Tyrosine Macrophage Colony-Stimulating Factor Phosphatidylinositol 3-Kinases Phosphotransferases (Alcohol Group Acceptor) Receptor Protein-Tyrosine Kinases Receptor, Macrophage Colony-Stimulating Factor
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
van der Geer P
Molecular Biology and Virology Laboratory, Salk Institute, San Diego, CA 92186-5800.
Hunter T
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1993-12-15
Pages
5161-72
Language
English
Region
England
NLM ID
8208664
PMCID
PMC413778
Subset
IM
Grants
NCI NIH HHS · CA14195 · United States
NCI NIH HHS · CA39780 · United States
External Links
PubMed source PMC Full Text View journal details
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