Browse > Article

v-Crk Induces Rac-dependent Membrane Ruffling and Cell Migration in CAS-deficient Embryonic Fibroblasts  

Sung, Bong Hwan (Center for Distributed Sensor Network (CDSN), Department of Life Sciences, Gwangju Institute of Science and Technology)
Yeo, Myoung Gu (Center for Distributed Sensor Network (CDSN), Department of Life Sciences, Gwangju Institute of Science and Technology)
Oh, Hye Jin (Center for Distributed Sensor Network (CDSN), Department of Life Sciences, Gwangju Institute of Science and Technology)
Song, Woo Keun (Center for Distributed Sensor Network (CDSN), Department of Life Sciences, Gwangju Institute of Science and Technology)
Abstract
Crk-associated substrate (CAS) is a focal adhesion protein that is involved in integrin signaling and cell migration. CAS deficiency reduces the migration and spreading of cells, both of which are processes mediated by Rac activation. We examined the functions of v-Crk, the oncogene product of the CT10 virus p47gag-crk, which affects cell migration and spreading, membrane ruffling, and Rac activation in CAS-deficient mouse embryonic fibroblasts (CAS-/- MEFs). CAS-/- MEFs showed less spreading than did CAS+/+ MEFs, but spreading was recovered in mutant cells that expressed v-Crk (CAS-/-v-Crk MEF). We observed that the reduction in spreading was linked to the formation of membrane ruffles, which were accompanied by Rac activation. In CAS-/- MEFs, Rac activity was significantly reduced, and Rac was not localized to the membrane. In contrast, Rac was active and localized to the membrane in CAS-/-v-Crk MEFs. Lamellipodia protrusion and ruffle retraction velocities were both reduced in CAS-/- MEFs, but not in CAS-/-v-Crk MEFs. We also found that microinjection of anti-gag antibodies inhibited the migration of CAS-/-v-Crk MEFs. These findings indicate that v-Crk controls cell migration and membrane dynamics by activating Rac in CAS-deficient MEFs.
Keywords
CAS; Cell Migration; Membrane Dynamics; Rac; v-Crk;
Citations & Related Records
Times Cited By KSCI : 1  (Citation Analysis)
Times Cited By Web Of Science : 3  (Related Records In Web of Science)
연도 인용수 순위
1 Lauffenburger, D.A. and Horwitz, A.F. (1996). Cell migration: a physically integrated molecular process. Cell 84, 359-369   DOI   ScienceOn
2 Rottner, K., Behrendt, B., Small, J.V., and Wehland, J. (1999). VASP dynamics during lamellipodia protrusion. Nat. Cell Biol. 1, 321-322   DOI   ScienceOn
3 Tsuda, M., Makino, Y., Iwahara, T., Nishihara, H., Sawa, H., Nagashima, K., Hanafusa, H., and Tanaka, S. (2004). Crk associates with ERM proteins and promotes cell motility toward hyaluronic acid. J. Biol. Chem. 279, 46843-46850   DOI   ScienceOn
4 Price, L.S., Leng, J., Schwartz, M.A., and Bokoch, G.M. (1998). Activation of Rac and Cdc42 by integrins mediates cell spreading. Mol. Biol. Cell 9, 1863-1871   DOI
5 Abassi, Y.A. and Vuori, K. (2002). Tyrosine 221 in Crk regulates adhesion-dependent membrane localization of Crk and Rac and activation of Rac signaling. EMBO J. 21, 4571-4582   DOI   ScienceOn
6 Wu, C.-C., Su, H.-W., Lee, C.-C., Tang, M.-J., and Su, F.-C. (2005). Quantitative measurement of changes in adhesion force involving focal adhesion kinase during cell attachment, spread, and migration. Biochem. Biophys. Res. Commun. 329, 256-265   DOI   ScienceOn
7 Matsuda, M., Mayer, B.J., Fukui, Y., and Hanafusa, H. (1990). Binding of transforming protein, P47gag-crk, to a broad range of phosphotyrosine-containing proteins. Science 248, 1537-1539   DOI
8 Nievers, M.G., Birge, R.B., Greulich, H., Verkleij, A.J., Hanafusa, H., and van Bergen en Henegouwen, P.M. (1997). v-Crk-induced cell transformation: changes in focal adhesion composition and signaling. J. Cell Sci. 110, 389-399
9 Stam, J.C., Geerts, W. J.C., Versteeg, H.H., Verkleij, A.J., and en Henegouwen, P. (2001). The v-Crk oncogene enhances cell survival and induces activation of protein kinase B/Akt. J. Biol. Chem. 276, 25176-25183   DOI   ScienceOn
10 Honda, H., Oda, H., Nakamoto, T., Honda, Z., Sakai, R., Suzuki, T., Saito, T., Nakamura, K., Nakao, K., Ishikawa, T., et al. (1998). Cardiovascular anomaly, impaired actin bundling and resistance to Src-induced transformation in mice lacking p130Cas. Nat. Genet. 19, 361-365   DOI   ScienceOn
11 Sugihara, K., Nakatsuji, N., Nakamura, K., Nakao, K., Hashimoto, R., Otani, H., Sakagami, H., Kondo, H., Nozawa, S., and Aiba, A. (1999). Rac 1 is required for the formation of three germ layers during gastrulation. Oncogene 17, 3427-3433   DOI
12 Kiyokawa, E., Hashimoto, Y., Kobayashi, S., Sugimura, H., Kurata, T., and Matsuda, M. (1998). Activation of Rac1 by a Crk SH3-binding protein, DOCK180. Genes Dev. 12, 3331-3336   DOI
13 Ridley, A.J. (2001). Rho GTPases and cell migration. J. Cell Sci. 114, 2713-2722
14 Yamada, K. M. and Geiger, B. (1997). Molecular interactions in cell adhesion complexes. Curr. Opin. Cell Biol. 9, 76-85   DOI   ScienceOn
15 Bear, J.E., Svitkina, T.M., Krause, M., Schafer, D.A., Loureiro, J.J., Strasser, G.A., Maly, I. V., Chaga, O.Y., Cooper, J.A., and Borisy, G.G. (2002). Antagonism between Ena/VASP proteins and actin filament capping regulates fibroblast motility. Cell 109, 509−521
16 Hinz, B., Alt, W., Johnen, C., Herzog, V., and Kaiser, H.W. (1999). Quantifying lamella dynamics of cultured cells by SACED, a new computer-assisted motion analysis. Exp. Cell. Res. 251, 234-243   DOI   ScienceOn
17 Lili, J., Yuchen, S., and Zhengtao, W. (2006). The Chemokine SDF-1alpha suppresses fibronectin-mediated in vitro lymphocytes adhesion. Mol. Cells 22, 308-313
18 Suetsugu, S., Yamazaki, D., Kurisu, S., and Takenawa, T. (2003). Differential roles of WAVE1 and WAVE2 in dorsal and peripheral ruffle formation for fibroblast cell migration. Dev. Cell 5, 595-609   DOI   ScienceOn
19 Yeo, M.G., Sung, B.H., Oh, H.J., Park, J.Y., Marcantonio, E.E., and Song, W.K. (2008). Focal adhesion targeting of v-Crk is essential for FAK phosphorylation and cell migration in mouse embryo fibroblasts deficient Src family kinases or p130CAS. J. Cell. Physiol. 214, 604-613   DOI   ScienceOn
20 del Pozo, M.A., Price, L.S., Alderson, N.B., Ren, X.D., and Schwartz, M.A. (2000). Adhesion to the extracellular matrix regulates the coupling of the small GTPase Rac to its effector PAK. EMBO J. 19, 2008-2014   DOI
21 Benard, V. and Bokoch, G.. (2002). Assay of CDC42, Rac, and Rho GTPase activation by affinity methods. In Methods in Enzymology, R. Iyengar and J.D. Hildebrandt, eds. (California USA: Academic Press), pp. 349-359
22 Allen, W.E., Jones, G.E., Pollard, J.W., and Ridley, A.J. (1997). Rho, Rac and Cdc42 regulate actin organization and cell adhesion in macrophages. J. Cell Sci. 110, 707-720
23 Mayer, B.J., Hamaguchi, M., and Hanafusa, H. (1988). A novel viral oncogene with structural similarity to phospholipase C. Nature 332, 272-275   DOI   ScienceOn
24 Pinco, K.A., He, W., and Yang, J.T. (2002). Alpha4beta1 integrin regulates lamellipodia protrusion via a focal complex/focal adhesion-independent mechanism. Mol. Biol. Cell 13, 3203-3217   DOI   ScienceOn
25 Tsuda, M., Tanaka, S., Sawa, H., Hanafusa, H., and Nagashima, K. (2002). Signaling adaptor protein v-Crk activates Rho and regulates cell motility in 3Y1 rat fibroblast cell line. Cell Growth Differ. 13, 131-139
26 Craig, S.W. and Chen, H. (2003). Lamellipodia protrusion: moving interactions of vinculin and Arp2/3. Curr. Biol. 13, 236-238   DOI   ScienceOn
27 Gu, J., Sumida, Y., Sanzen, N., and Sekiguchi, K. (2001). Laminin-10/11 and fibronectin differentially regulate integrin-dependent Rho and Rac activation via p130Cas-CrkII-DOCK180 pathway. J. Biol. Chem. 276, 27090-27097   DOI   ScienceOn
28 Honda, H., Nakamoto, T., Sakai, R., and Hirai, H. (1999). p130Cas, an assembling molecule of actin filaments, promotes cell movement, cell migration, and cell spreading in fibroblasts. Biochem. Biophys. Res. Commun. 262, 25-30   DOI   ScienceOn
29 Ridley, A.J., Paterson, H.F., Johnston, C.L., Diekmann, D., and Hall, A. (1992). The small GTP-binding protein rac regulates growth factor-induced membrane ruffling. Cell 70, 401-410   DOI   ScienceOn
30 Huttenlocher, A., Sandborg, R.R., and Horwitz, A.F. (1995). Adhesion in cell migration. Curr. Opin. Cell. Biol. 7, 697-706   DOI   ScienceOn
31 Klemke, R.L., Leng, J., Molander, R., Brooks, P.C., Vuori, K., and Cheresh, D.A. (1998). CAS/Crk coupling serves as a "molecular switch" for induction of cell migration. J. Cell Biol. 140, 961-972   DOI   ScienceOn
32 Kanner, S.B., Reynolds, A.B., Wang, H. C., Vines, R.R., and Parsons, J.T. (1991). The SH2 and SH3 domains of pp60src direct stable association with tyrosine phosphorylated proteins p130 and p110. EMBO J. 10, 1689-1698
33 Altun-Gultekin, Z.F., Chandriani, S., Bougeret, C., Ishizaki, T., Narumiya, S., de Graaf, P., Van Bergen en Henegouwen, P., Hanafusa, H., Wagner, J.A., and Birge, R.B. (1998). Activation of Rho-dependent cell spreading and focal adhesion biogenesis by the v-Crk adaptor protein. Mol. Cell. Biol. 18, 3044-3058   DOI
34 Legg, J.A., Bompard, G., Dawson, J., Morris, H.L., Andrew, N., Cooper, L., Johnston, S.A., Tramountanis, G., and Machesky, L.M. (2007). N-WASP involvement in dorsal ruffle formation in mouse embryonic fibroblasts. Mol. Biol. Cell 18, 678-687   DOI   ScienceOn
35 Bear, J.E., Loureiro, J.J., Libova, I., Faesler, R., Wehland, J., and Gertler, F.B. (2000). Negative regulation of fibroblast motility by Ena/VASP proteins. Cell 101, 717-728   DOI   ScienceOn