Summary
Upon cell adhesion to extracellular matrix proteins, focal adhesion kinase (FAK) rapidly undergoes autophosphorylation on its Tyr-397 which consequently serves as a binding site for the Src homology 2 domains of the Src family protein kinases and several other intracellular signaling molecules. In this study, we have attempted to examine the effect of the FAK Y397F mutant on v-Src-stimulated cell transformation by establishing an inducible expression of the Y397F mutant in v-Src-transformed FAK-null (FAK−/−) mouse embryo fibroblasts. We found that the FAK Y397F mutant had both positive and negative effects on v-Src-stimulated cell transformation; it promoted v-Src-stimulated invasion, but on the other hand it inhibited the v-Src-stimulated anchorage-independent cell growth in vitro and tumor formation in vivo . The positive effect of the Y397F mutant on v-Src-stimulated invasion was correlated with an increased expression of matrix metalloproteinase-2, both of which were inhibited by the specific phosphatidylinositol 3-kinase inhibitor wortmannin or a dominant negative mutant of AKT, suggesting a critical role for the phosphatidylinositol 3-kinase/AKT pathway in both events. However, the expression of the Y397F mutant rendered v-Src-transformed FAK−/− cells susceptible to anoikis, correlated with suppression on v-Src-stimulated activation of ERK and AKT. In addition, under anoikis stress, the induction of the Y397F mutant in v-Src-transformed FAK−/− cells selectively led to a decrease in the level of p130Cas, but not other focal adhesion proteins such as talin, vinculin, and paxillin. These results suggest that FAK may increase the susceptibility of v-Src-transformed cells to anoikis by modulating the level of p130Cas.
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Abbreviations
- Cas:
-
Crk-associated substrate
- ERK:
-
extracellular signal-regulated kinase
- FAK:
-
focal adhesion kianse
- HA:
-
hemagglutinin
- IB:
-
immunoblotting
- IP:
-
immunoprecipitation
- JNK:
-
c-Jun N-terminal kinase
- MMP:
-
matrix metalloproteinase
- PI3K:
-
phosphatidylinositol 3-kinase
- SH:
-
Src homology
- Src:
-
sarcoma
- STAT3:
-
signal transducers and activators of transcription 3
- Tet:
-
tetracycline
- Tyr:
-
tyrosine
- v-Scr:
-
viral Scr
- Wt:
-
wild type
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Chang, LC., Huang, CH., Cheng, CH. et al. Differential Effect of the Focal Adhesion Kinase Y397F Mutant on v-Src-Stimulated Cell Invasion and Tumor Growth. J Biomed Sci 12, 571–585 (2005). https://doi.org/10.1007/s11373-005-7212-5
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DOI: https://doi.org/10.1007/s11373-005-7212-5