Differential regulation of cell migration, actin stress fiber organization, and cell transformation by functional domains of Crk-associated substrate.
The Crk-associated substrate (Cas) is a unique docking protein that possesses a repetitive stretch of tyrosine-containing motifs and an Src homology 3 (SH3) domain. Embryonic fibroblasts lacking Cas demonstrated resistance to Src-induced transformation along with impaired actin bundling and cell motility, indicating critical roles of Cas in actin cytoskeleton organization, cell migration, and oncogenesis. To gain further insight into roles of each domain of Cas in these processes, a compensation assay was performed by expressing a series of Cas mutants in Cas-deficient fibroblasts. The results showed that motifs containing YDxP were indispensable for actin cytoskeleton organization and cell migration, suggesting that CrkII-mediated signaling regulates these biological processes. The C-terminal Src-binding domain played essential roles in cell migration and membrane localization of Cas, although it was dispensable in the organization of actin stress fibers. Furthermore, the Src-binding domain was also a prerequisite for Src transformation possibly, because of its crucial role in the phosphorylation of Cas during transformation. Overall, differential uses of the Cas domains in individual biological processes were demonstrated.
Huang
Jinhong
J
Cancer Signal Transduction Project, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045.
Hamasaki
Hiroko
H
Nakamoto
Tetsuya
T
Honda
Hiroaki
H
Hirai
Hisamaru
H
Saito
Masaki
M
Takato
Tsuyoshi
T
Sakai
Ryuichi
R
D000199:Actins / Q000378:metabolism*; D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D002465:Cell Movement; D003599:Cytoskeleton; D018076:DNA, Complementary / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D015151:Immunoblotting; D008856:Microscopy, Fluorescence; D008957:Models, Genetic; D009154:Mutation; D010750:Phosphoproteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism*; D010766:Phosphorylation; D011233:Precipitin Tests; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D051381:Rats; D015398:Signal Transduction; D018909:src Homology Domains
D000199:Actins; D048868:Adaptor Proteins, Signal Transducing; D018076:DNA, Complementary; C099942:NEDD9 protein, human; D010750:Phosphoproteins