C3G is tyrosine-phosphorylated after integrin-mediated cell adhesion in normal but not in Bcr/Abl expressing cells.
The SH2-SH3 adaptor protein Crkl has been implicated in the signal transduction pathways of several membrane-bound receptors. Tyrosine phosphorylation of proteins associated with such signalling complexes can generate binding sites for the Crkl SH2-domain and can recruit proteins constitutively bound to Crkl via the Crkl SH3 domain into such complexes. In the current study we show that Crkl, but only a minor amount of the related Crk, form constitutive complexes in vivo with guanine nucleotide exchange factor C3G in 3T3 fibroblasts. Adhesion of both normal and transformed cells to fibronectin or other extracellular matrix proteins consistently induces the tyrosine-phosphorylation of C3G. Adhesion-induced tyrosine phosphorylation of C3G is dependent on an intact cytoskeleton and peaks at 5-10 min after attachment. In contrast, 3T3 cells stably transfected with Bcr/Abl P210 show a prominent reduction in the amount of C3G complexed to Crkl and do not exhibit tyrosine-phosphorylation of C3G upon spreading and attachment. These data establish that integrin-mediated cell adhesion results in Crkl-mediated tyrosine phosphorylation of C3G, a pathway which can be disrupted by Bcr/Abl.
de Jong
R
R
Department of Pathology, Childrens Hospital Los Angeles Research Institute and USC School of Medicine, California 90027, USA.
van Wijk
A
A
Heisterkamp
N
N
Groffen
J
J
D016475:3T3 Cells; D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D001665:Binding Sites; D002448:Cell Adhesion; D003599:Cytoskeleton / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism; D016044:Fusion Proteins, bcr-abl / Q000502:physiology*; D020662:Guanine Nucleotide Exchange Factors; D016023:Integrins / Q000502:physiology*; D046911:Macromolecular Substances; D051379:Mice; D009687:Nuclear Proteins / Q000378:metabolism*; D010766:Phosphorylation; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000502:physiology; D015398:Signal Transduction; D014162:Transfection; D018909:src Homology Domains
D048868:Adaptor Proteins, Signal Transducing; C082869:CRKL protein; D016326:Extracellular Matrix Proteins; D005353:Fibronectins; D020662:Guanine Nucleotide Exchange Factors; D016023:Integrins; D046911:Macromolecular Substances; D009687:Nuclear Proteins; D011506:Proteins; D011993:Recombinant Fusion Proteins; D016044:Fusion Proteins, bcr-abl