Evidence that DOCK180 up-regulates signals from the CrkII-p130(Cas) complex.
DOCK180 is one of the two principal proteins bound to the SH3 domain of the adaptor protein CrkII. Here, we have studied the involvement of DOCK180 in integrin signaling. DOCK180 was neither phosphorylated nor bound to CrkII in quiescent NIH 3T3 cells and 3Y1 cells. We found that DOCK180 was phosphorylated and bound to CrkII in NIH 3T3 cells stimulated with integrin and also in 3Y1 cells transformed by v-src or v-crk. The binding of DOCK180 to CrkII correlated with the binding of CrkII to p130(Cas), which is a major CrkII SH2 domain-binding protein at focal adhesions. In a reconstitution experiment, expression of DOCK180 induced hyperphosphorylation of p130(Cas) and a concomitant increase in the amount of CrkII bound to p130(Cas). Similarly, binding of DOCK180 to CrkII was also enhanced by the coexpression of p130(Cas). Finally, we found that coexpression of p130(Cas) and CrkII with DOCK180 induced local membrane spreading and accumulation of DOCK180-CrkII-p130(Cas) complexes at focal adhesions. These findings suggest that DOCK180 positively regulates signaling from integrins to CrkII-p130(Cas) complexes at focal adhesions.
Kiyokawa
E
E
Department of Pathology, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8840, Japan.
Hashimoto
Y
Y
Kurata
T
T
Sugimura
H
H
Matsuda
M
M
D016475:3T3 Cells; D000818:Animals; D002460:Cell Line; D002461:Cell Line, Transformed; D003001:Cloning, Molecular; D050719:Crk-Associated Substrate Protein; D004815:Epidermal Growth Factor / Q000494:pharmacology / Q000502:physiology; D005353:Fibronectins / Q000494:pharmacology; D016391:Genes, src; D006801:Humans; D016023:Integrins / Q000502:physiology; D007668:Kidney; D051379:Mice; D050736:Oncogene Protein v-crk; D009857:Oncogenes; D010750:Phosphoproteins / Q000302:isolation & purification / Q000378:metabolism*; D010766:Phosphorylation; D011107:Polylysine / Q000494:pharmacology; D011485:Protein Binding; D011494:Protein Kinases / Q000302:isolation & purification / Q000378:metabolism*; D011506:Proteins / Q000302:isolation & purification / Q000378:metabolism*; D011518:Proto-Oncogene Proteins; D050737:Proto-Oncogene Proteins c-crk; D011994:Recombinant Proteins / Q000378:metabolism; D050717:Retinoblastoma-Like Protein p130; D015682:Retroviridae Proteins, Oncogenic / Q000235:genetics; D015398:Signal Transduction; D014162:Transfection; D020744:rac GTP-Binding Proteins; D018909:src Homology Domains
C493841:BCAR1 protein, human; C493842:Bcar1 protein, mouse; D050719:Crk-Associated Substrate Protein; C099818:DOCK1 protein, human; D005353:Fibronectins; D016023:Integrins; D050736:Oncogene Protein v-crk; D010750:Phosphoproteins; D011506:Proteins; D011518:Proto-Oncogene Proteins; D050737:Proto-Oncogene Proteins c-crk; D011994:Recombinant Proteins; D050717:Retinoblastoma-Like Protein p130; D015682:Retroviridae Proteins, Oncogenic; D011107:Polylysine; D004815:Epidermal Growth Factor; D011494:Protein Kinases; D020744:rac GTP-Binding Proteins