Signaling adaptor protein v-Crk activates Rho and regulates cell motility in 3Y1 rat fibroblast cell line.
The adaptor protein Crk has been reported to associate with focal adhesions and is thought to be involved in integrin-mediated signaling pathway. However, the precise mechanism of Crk-dependent regulation of cytoskeleton still remains under investigation. In this study, we have established a v-Crk-inducible cell line in rat fibroblasts 3Y1 cells and found that v-Crk activated Rho and induced actin stress fiber formation. In addition to the induction of tyrosine-phosphorylation of p130(Cas) and paxillin, we demonstrated that v-Crk induced threonine-phosphorylated bands sized at 72/78 kDa found specifically in 3Y1 cells. Both of the inhibitors of Rho and Rho-associated kinase, C3 and Y27632, respectively, inhibited these v-Crk-induced biochemical effects. Although v-Crk-induced cells exhibited a decrease of cell motility, integrin stimulation recovered the suppression of motility. Furthermore, v-Crk enhanced motility in chemotactic assay toward fibronectin with additional activation of Rho and the increase of levels of CD44 cleavage. These results suggest that v-Crk activated Rho and induced actin stress fiber formation and CD44 cleavage leading to the regulation of cell motility.
Tsuda
Masumi
M
Laboratory of Molecular and Cellular Pathology, Hokkaido University School of Medicine, Sapporo 060-8638, Japan.
Tanaka
Shinya
S
Sawa
Hirofumi
H
Hanafusa
Hidesaburo
H
Nagashima
Kazuo
K
D000209:Acute-Phase Proteins / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D002465:Cell Movement; D002478:Cells, Cultured; D005347:Fibroblasts / Q000378:metabolism*; D050736:Oncogene Protein v-crk; D010766:Phosphorylation; D010769:Phosphothreonine / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D015682:Retroviridae Proteins, Oncogenic / Q000235:genetics / Q000494:pharmacology*; D015398:Signal Transduction; D013912:Threonine / Q000378:metabolism
D000209:Acute-Phase Proteins; D050736:Oncogene Protein v-crk; D015682:Retroviridae Proteins, Oncogenic; C010781:acute-phase protein rho; D010769:Phosphothreonine; D013912:Threonine