Untersuchungen zum Rekrutierungsmechanismus und zur funktionellen Rolle des atypischen Myr5 bei der Epithelzellinvasion durch Shigella flexneri Christian Böwe 1.51.1.61.2.7891.3.1011605#38712132.142.1.2.2.2.3.152.4.162.5.2.6.172.6.1.2.6.2.182.7.2.7.1.19202.7.2.2.8.2.9.2.9.1.212.9.2.2.9.3.222.9.4.2.9.5.232.10.242.10.1.2.10.2.252.10.3.262.10.4.272.10.5.282.10.6.293.303.1.3.2.3132333.3.343.3.1.35363.3.2.373.4.3839403.5.41423.6.43604#81544453.7.464748590#3533.8.4950617#3724.514.1.4.1.1.4.1.2.524.1.3.4.1.4.534.1.5.4.2.545556574.3.58596061626364655.6667 6. Literaturverzeichnis68697071727374757677 Abkürzungsverzeichnis7879 Lebenslauf80 Danksagung81 Selbständigkeitserklärung82deInhaltsverzeichnisHilfe 6. Literaturverzeichnis (1) Sansonetti, P.J. 1991. Genetic and molecular basis of epithelial cell invasion by Shigella species. Rev. Infect. Dis. 13 : 285-292 (2) Ginocchio, C.C., Olmsted, S.B., Wells, C.L., Galan, J.E. 1994. Contact with epithelial cells induces the formation of surface appendages on Salmonella typhimurium. Cell 76 : 717-724 (3) Hardt, W.D., Chen, L.M., Schuebel, K.E., Bustelo, X.R., Galan, J.E. 1998. S. typhimurium encodes an activator of Rho GTPases that induces membrane ruffling and nuclear responses in host cells. Cell 93 : 815-826 (4) Falkow, S., Isberg, R.R., Portnoy, D.A. 1992. The interaction of bacteria with mammalian cells. Annu. Rev. Cell. Biol. 8 : 333-363 (5) Finlay, B.B., Falkow, S. 1989. Common themes in microbial pathogenicity. Microbiol. Rev. 53 : 476-511 (6) Sanders, M.C., Theriot, J.A. 1996. Tails from the hall of infection: actin-based motility of pathogens. Trends Microbiol . 4 : 211-213 (7) Winkler, H.H. 1990. Rickettsia species (as organism). Annu. Rev. Microbiol . 44 : 131-133 (8) Gailard, J.L., Berche, P., Mounier, J., Richard, S., Sansonetti, P. 1987. In vitro model of penetration and intracellular growth of Listeria monocytogenes in the human enterocyte cell line Caco2. Infect. Immun . 55 : 2822-2829 (9) Menard, R., Dehio, C., Sansonetti, P.J. 1996. Bacterial entry into epithelial cells: the paradigm of Shigella . Trends Microbiol. 4 : 220-226 (10) High, N., Mounier, J., Prevost, M.C., Sansonetti, P.J. 1992. IpaB of Shigella flexneri causes entry into epithelial cells and escape from the phagocytic vacuole. EMBO J. 11 : 1991-1999 (11) Loisel, T.P., Boujemaa, R., Pantaloni, D., Carlier, M.F. 1999. Reconstitution of actin-based motility of Listeria and Shigella using pure proteins. Nature 401 : 613-616 (12) Cossart, P., Lecuit, M. 1998. Interactions of Listeria monocytogenes with mammalian cells during entry and actin-based movement: bacterial factors, cellular ligands and signaling. EMBO J. 17 : 3797-3806 (13) Niebuhr, K. 1994. Untersuchungen zu Pathogenitätsmechanismen von Listeria monocytogenes: Charakterisierung des bakteriellen Virulenzfaktors ActA. Disser­tation,

TU Braunschweig

(14) Egile, C., Loisel, T.P., Laurent, V., Pantaloni, D., Sansonetti, P.J., Carlier, M.F. 1999. Activation of the CDC42 effector N-WASP by the Shigella flexneri IcsA protein promotes actin nucleation by Arp2/3 complex and bacterial actin-based motility. J. Cell Biol. 146 : 1319-1332 (15) Collier, L., Balows, A., Sussman, M. 1998. Topley and Wilson`s microbiology and microbial infections / ninth edition, volume 2, systemic bacteriology. Oxford University Press, Inc.,

New York:

943-945 (16) Pupo, G.M., Karaolis, D.K., Lan, R., Reeves, P.R. 1997. Evolutionary relationship among pathogenic and nonpathogenic Escherichia coli strains inferred from multilocus enzyme electrophoresis and mdh sequence studies. Infect.Immun. 65 : 2685-2692 (17) Pupo, G.M., Lan, R., Reeves, P.R. 2000. Multiple independent origins of Shigella clones of Escherichia coli and convergent evolution of many of their characteristics. Proc. Natl. Acad. Sci.USA 97 : 10567-10572 (18) Rolland, K., Lambert, Zechovsky, N., Picard, B., Denamur, E. 1998. Shigella and enteroinvasive Escherichia coli strains are derived from distinct ancestral strains of E.coli. Microbiology 144 : 2667-2672 (19) Hornick, R.B. 1978. Bacterial infections of the intestine. aus: Weinstein, L., Fields, B.N. Seminars in infectious disease, chap 3. Stratton Intercontinental Medical,

New York

: 68-96 (20) Brandis, H., Eggers, H.J., Köhler, W., Pulverer, G. 1994. Lehrbuch der Medizinischen Mikrobiologie , 7. Auflage. Gustav Fischer Verlag⋅

Stuttgart

⋅

Jena

⋅

New York

: 404-408 (21) Perdomo, J.J., Cavaillon, J.M., Huerre, M., Ohayon, H., Gounon, P., Sansonetti, P.J. 1994. Acute inflammation causes epithelial invasion and mucosal destruction in experimental shigellosis. J. Exp. Med. 180 : 1307-1319 (22) Clerc, P., Sansonetti, P.J. 1987. Entry of Shigella flexneri into HeLa cells: evidence for directed phagocytosis involving actin polymerization and myosin accumulation. Infect. Immun. 55 : 2681-2688 (23) Neutra, M.R., Frey, A., Kraehenbuhl, J.P. 1996. Epithelial M cells: gateways for mucosal infection and immunization. Cell 86 : 345-348 (24) Zychlinsky, A., Prevost, M.C., Sansonetti, P.J. 1992. Shigella flexneri induces apoptosis in infected macrophages. Nature 358 : 167-169 (25) Zychlinsky, A., Fitting, C., Cavaillon, J.M., Sansonetti, P.J. 1994. Interleukin 1 is released by murine macrophages during apoptosis induced by Shigella flexneri . J. Clin. Invest. 94 : 1328-1332 (26) Sansonetti, P.J., Arondel, J., Cavaillon, J.M., Huerre, M. 1995. Role of interleukin-1 in the pathogenesis of experimental shigellosis. J. Clin. Invest. 96 : 884-892 (27) Mounier, J., Vasselon, T., Hellio, R., Lesourd, M., Sansonetti, P.J. 1992. Shigella flexneri enters human colonic Caco-2 epithelial cells through the basolateral pole. Infect. Immun. 60 : 237-248 (28) Perdomo, J.J., Gounon, P., Sansonetti, P.J. 1994. Polymorphonuclear leucocyte transmigration promotes invasion of colonic epithelial monolayers by Shigella flexneri. J. Clin. Invest. 93 : 633-643 (29) Beatty, W.L., Sansonetti, P.J. 1997. Role of lipopolysaccharide in signalling to subepithelial polymorphonuclear leucocytes. Infect. Immun. 65 : 4395-4404 (30) Goldberg, M.B., Sansonetti, P.J. 1993. Shigella subversion of the cellular cytoskeleton: a strategy for epithelial colonization. Infect. Immun. 61 : 4941-4946 (31) Allaoui, A., Mounier, J., Prevost, M.C., Sansonetti, P.J., Parsot, C. 1992. IcsB: a Shigella flexneri virulence gene necessary for the lysis of protrusions during intercellular spread. Mol. Microbiol. 6 : 1605-1616 (32) Goldberg, M.B., Bazu, O., Parsot, C., Sansonetti, P.J. 1993. Unipolar localization and ATPase activity of IcsA, a Shigella flexneri protein involved in intracellular movement. J. Bacteriol. 175 : 2189-2196 (33) Bernardini, M.L., Mounier, J., d´Hauteville, H., Coquis-Rondon, M., Sansonetti, P.J. 1989. Identification of icsA, a plasmid locus of Shigella flexneri which governs bacterial intra- and intercellular spread through interaction with F-actin. Proc. Natl. Acad. Sci. USA 86 : 3867-3871 (34) Sansonetti, P.J., Arondel, J., Fontaine, A., d´Hauteville, H., Bernardini, M.L. 1991. OmpB (osmo-regulation) and icsA (cell-to-cell-spread) mutants of Shigella flexneri : vaccine candidates and probes to study the pathogenesis of shigellosis. Vaccine 9 : 416-422 (35) Vasselon, T., Mounier, J., Prevost, M.C., Hellio, R., Sansonetti, P.J. 1991. Stress fiber-based movement of Shigella flexneri within cells. Infect. Immun. 59 : 1723-1732 (36) Vasselon, T., Mounier, J., Hellio, R., Sansonetti, P.J. 1992. Movement along actin filaments of the perijunctional area and de novo polymerization of cellular actin are required for Shigella flexneri colonization of epithelial Caco-2 cell monolayers. Infect. Immun. 60 : 1031-1040 (37) Adam, T., Giry, M., Boquet, P., Sansonetti, P. 1996. Rho-dependent membrane folding causes Shigella entry into epithelial cells. EMBO J. 15 : 3315-3321 (38) D´Hauteville, H., Lagelouse, R.D., Nato, F., Sansonetti, P.J. 1996. Lack of cleavage of IcsA in Shigella flexneri causes aberrant movement and allows demonstration of a cross-reactive eucaryotic protein. Infect. Immun. 64 : 511-517 (39) Labrec, E.H., Schneider, H., Magnani, T.J., Formal, S.B. 1964. Epithelial cell penetration as an essential step in the pathogenesis of bacillary dysentery. J. Bact. 88 : 1503-1518 (40) Sansonetti, P.J., Arondel, J., Cantey, J.R., Prevost, M.C., Huerre, M. 1996. Infection of rabbit Peyer´s patches by Shigella flexneri : effect of adhesive or invasive bacterial phenotypes on follicle-associated epithelium. Infect. Immun. 64 : 2752-2764 (41) Adam, T., Arpin, M., Prevost, M.C., Gounon, P., Sansonetti, P.J. 1995. Cytoskeletal rearrangements and the functional role of T-Plastin during entry of Shigella flexneri into HeLa cells. J. Cell Biol. 129 : 367-381 (42) Watarai, M., Tobe, T., Yoshikawa, M., Sasakawa, C. 1995. Contact of Shigella with host cells triggers release of Ipa invasins and is an essential function of invasiveness. EMBO J. 14 : 2461-2470 (43) Barzu, S., Benjelloun-Touimi, Z., Phalipon, A., Sansonetti, P., Parsot, C. 1997. Functional analysis of the Shigella flexneri IpaC invasin by insertional mutagenesis. Infect. Immun. 65 : 1599-1605 (44) Menard, R., Prevost, M.C., Gounon, P., Sansonetti, P., Dehio, C. 1996. The secreted Ipa complex of Shigella flexneri promotes entry into mammalian cells. Proc. Natl. Acad. Sci. USA 93 : 1254-1258 (45) Tran Van Nhieu, G., Ben-Ze´ev, A., Sansonetti, P.J. 1997. Modulation of bacterial entry into epithelial cells by association between vinculin and the Shigella IpaA invasin. EMBO J. 16 : 2717-2729 (46) Menard, R., Sansonetti, P., Parsot, C. 1994. The secretion of the Shigella flexneri Ipa invasins is activated by epithelial cells and controlled by IpaB and IpaD. EMBO J. 13 : 5293-5302 (47) Menard, R., Sansonetti, P., Parsot, C., Vasselon, T. 1994. Extracellular association and cytoplasmic partitioning of the IpaB and IpaC invasins of S. flexneri . Cell 79: 515-525 (48) Tran van Nhieu, G., Caron, E., Hall, A., Sansonetti, P.J. 1999. IpaC induces actin polymerization and filopodia formation during Shigella entry into epithelial cells. Embo J. 18 : 3249-3262 (49) Mounier, J., Bahrani, F.K., Sansonetti, P.J. 1997. Secretion of Shigella flexneri Ipa invasins on contact with epithelial cells and subsequent entry of the bacterium into cells are growth stage dependent. Infect. Immun. 65 : 774-782 (50) Bourdet-Sicard, R., Rüdiger, M., Jockusch, B.M., Gounon, P., Sansonetti, P.J., Tran Van Nhieu, G. 1999. Binding of the Shigella protein IpaA to vinculin induces F-actin depolymerization. Embo J. 18 : 5853-5862 (51) Halle, T.L., Morris, R.E., Bonventre, P.F. 1979. Shigella infection of Henle intestinal epithelial cells: role of the host cell. Infect. Immun. 24 : 887-894 (52) Skoudy, A., Tran Van Nhieu, G., Mantis, N., Arpin, M., Mounier, J., Gounon, P., Sansonetti, P. 1999. A functional role for ezrin during Shigella flexneri entry into epithelial cells. J. Cell Sci. 112 : 2059-2068 (53) Mounier, J., Laurent, V., Hall, A., Fort, P., Carlier, M.F., Sansonetti, P.J., Egile, C. 1999. Rho family GTPases control entry of Shigella flexneri into epithelial cells but not intracellular motility . J. Cell Sci. 112 : 2069-2080 (54) Amano, M., Chihara, K., Kimura, K., Fukata, Y., Nakamura, N., Matsuura, Y., Kaibuchi, K. 1997. Formation of actin stress fibers and focal adhesions enhanced by Rho-kinase. Science 275 : 1308-1311 (55) Allen, W.E., Gareth, E.J., Jeffrey, W.P., Ridley, A.J. 1997. Rho, Rac and CDC42 regulate actin organization and cell adhesion in macrophages. J. Cell Sci. 110 : 707-720 (56) Ridley, A.J., Hall, A. 1992. The small GTP-binding protein Rho regulates the assembly of focal adhesions and stress fibers in response to growth factors. Cell 70 : 389-399 (57) Bowman, E.P., Uhlinger, D.J., Lambeth, J.D. 1993. Neutrophil phospholipase D is activated by a membrane-associated rho family small molecular weight GTP-binding protein. J. Biol. Chem. 268 : 24535-24538 (58) Chong, L.D., Traynor-Kaplan, A., Bokoch, G.M., Schwartz, M.A. 1994. The small GTP-binding protein rho regulates a phosphatidylinositol 4-phosphate 5-kinase in mammalian cells. Cell 79 : 507-513 (59) Watari, M., Kamata, Y., Kozaki, S., Sasakawa, C. 1997. Rho, a small GTP-binding protein, is essential for Shigella invasion of epithelial cells. J. Exp. Med. 185 : 281-292 (60) Narumiya, S., Morii, N. 1993. Rho gene products, botulinum C3 exoenzym and cell adhesion. Cell. Sign. 5 : 9-19 (61) Machesky, L.M., Hall, A. 1997. Role of actin polymerization and adhesion to extra-cellular matrix in Rac- and Rho-induced cytoskeletal reorganization. J. Cell Biol. 138 : 913-926 (62) Wu, W.J., Leonard, D.A., Cerione, A.R., Manor, D. 1997. Interaction between CDC42 and RhoGDI is mediated through the Rho insert region. J. Biol. Chem. 272 : 26153-26158 (63) Reinhard, J., Scheel, A.A., Diekmann, D., Hall, A., Ruppert, C., Bähler, M. 1995. A novel type of myosin implicated in signalling by Rho family GTPases. EMBO J. 14 : 697-704 (64) Thompson, R.F., Langford, G.M., 2002. Myosin superfamily evolutionary history. Anat Rec 268 : 276-289 (65) Mermall, V., Post, P.L., Mooseker, M.S. 1998. Unconventional myosins in cell movement, membrane traffic, and signal transduction. Science 279 : 527-533 (66) Hasson, T., Mooseker, S. 1995. Molecular motors, membrane movements and physiology: emerging roles for myosins. Curr. Opin. Cell Biol. 7 : 587-594 (67) Titus, M.A. 1997. Unconventional myosins: new frontiers in actin-based motors. Trends Cell Biol. 7 : 119-123 (68) Voigt, H., Olivo, J.C., Sansonetti, P., Guillen, N. 1999. Myosin IB from Entamoeba histolytica is involved in phagocytosis of human erythrocytes. J. Cell Sci. 112 : 1191-1201 (69) Self, T., Mahony, M., Fleming, J., Walsh, J., Brown, S.D.M., Steel, K.P. 1998. Shaker-1 mutations reveal roles for myosin VIIA in both development and function of cochlear hair cells. Development 125 : 557-566 (70) Wirth, J.A., Jensen, K.A., Post, P.L., Bement, W.M., Mooseker, M.S. 1996. Human myosin-IXb, an unconventional myosin with a chimerin-like rho/rac GTPase-activating protein domain in its tail. J. Cell Sci. 109 : 653-661 (71) Post, P.L., Bokoch, G.M., Mooseker, M.S. 1998. Human myosin-IXb is a mechano-chemically active motor and a GAP for rho . J. Cell Sci. 111 : 941-950 (72) Chieregatti, E., Gärtner, A., Stöffler, H.E., Bähler, M. 1998. Myr 7 is a novel myosin IX-RhoGAP expressed in rat brain. J. Cell Sci. 111 : 3597-3608 (73) Müller, R.T., Honnert, U., Reinhard, J., Bähler, M. 1997. The rat myosin myr 5 is a GTPase-activating protein for rho in vivo: essential role of arginine 1695. Mol. Biol. Cell 8 : 2039-2053 (74) Kreis, T.E. 1986. Microinjected antibodies against the cytoplasmic domain of vesicular stomatis virus glycoprotein block its transport to the cell surface. EMBO J. 5 : 931-941 (75) Maurelli, A.T., Baudry, B., d´Hauteville, H., Sansonetti, P.J. 1985. Cloning of plasmid DNA sequences involved in invasion of HeLa cells by Shigella flexner i. Infect. Immun. 49 : 164-171 (76) Sansonetti, P.J., Ryter, A., Clerc, P., Maurelli, A.T., Mounier, J. 1986. Multiplication of Shigella flexneri within HeLa cells: lysis of the phagocytic vacuole and plasmid-mediated contact hemolysis. Infect. Immun. 51 : 461-469 (77) Labigne-Roussel, A.F., Lark, D., Schoolnik, G., Falkow, S. 1984. Cloning and expression of an afimbrial adhesin (AFA-I) responsible for P blood group-independent, mannose-resistant hemagglutination from a pyelonephritic Escherichia coli strain. Infect. Immun. 46 : 251-259 (78) Hsu, S.H., Schacter, B.Z., Delaney, N.L., Miller, T.B., McKusick, V.A., Kennett, R.H., Bodmer, J.G., Young, D., Bodmer, W.F. 1976. Genetic characteristics of the HeLa cell. Science 191 : 1966-1982 (79) Alberts, B., Bray, D., Lewis, J., Raff, M., Roberts, K., Watson, J.D. 1994. Molecular Biology of the cell / Third Edition. Garland Publishung, Inc.,

New York-London

: 847-858 (80) Bejsovec, A., Anderson, P. 1990. Function of the myosin ATP and actin binding sites are required for C. elegans thick filament assembly. Cell 60 : 133-140 (81) Shimada, T., Sasaki, N., Ohkura, R., Sutoh, K. 1997. Alanine scanning mutagenesis of the switch I region in the ATPase site of Dictyostelium discoideum myosin II. Biochemistry 36 : 14037-14043 (82) Sansonetti, P.J. 1992. Molecular and cellular biology of Shigella flexneri invasiveness: from cell assay systems to shigellosis. aus: Current Topics in Microbiology and Immunology, Vol. 180, Springer-Verlag Berlin-Heidelberg: 1-18 (83) Rousset, M. 1986. The human colon carcinoma cell lines HT-29 and Caco-2: two in vitro models for the study of intestinal differentiation. Biochemie . 68 : 1035-1400 (84)Mermall, V., McNally, J.G., Miller, K.G. 1994. Transport of cytoplasmic particles catalysed by an unconventional myosin in living Drosophila embryos. Nature 369 : 560-562 (85) Govindan, B., Bowser, R., Novick, P. 1995. The role of Myo2, a yeast class V myosin, in vesicular transport . J. Cell Biol. 128 : 1055-1068 (86) Tabb, J.S., Molyneaux, B.J., Cohen, D.L., Kuznetsov, S.A., Langford, G.M. 1998. Transport of ER vesicles on actin filaments in neurons by myosin V. J. Cell Sci. 111 : 3221-3234 (87) Evans, L.L., Lee, A.J., Bridgman, P.C., Mooseker, M.S. 1998. Vesicle-associated brain myosin-V can be activated to catalyze actin-based transport. J. Cell Sci. 111 : 2055-2066 (88) Haarer, B.K., Petzold, A., Lillie, S.H., Brown, S.S. 1994. Identification of MYO4, a second class V myosin gene in yeast. J. Cell Sci. 107 : 1055-1064 (89) Hasson, T.B., Mooseker, M.S. 1992. Two novel unconventional myosins identified from a kidney proximal tubule cell line. Mol. Biol. Cell 3 : 158 (90) Hammer, J.A. III. 1994. The structure and function of unconventional myosins: a review. J. Muscle Res. Cell Motil. 15 : 1-10 (91) Alberts, B., Bray, D., Lewis, J., Raff, M., Roberts, K., Watson, J.D. 1994. Molecular Biology of the cell / Third Edition. Garland Publishung, Inc.,

New York-London

: 834-847 (92) Alberts, B., Bray, D., Lewis, J., Raff, M., Roberts, K., Watson, J.D. 1994. Molecular biology of the cell / Third Edition. Garland Publishung, Inc.,

New York-London

: 813-814 (93) Fath, K.R., Trimbur, G.M., Burgess, D.R. 1994. Molecular motors are differentially distributed on golgi membranes from polarized epithelial cells. J. Cell Biol. 126 : 661-675 (94) Alcalde, J., Egea, G., Sandoval, I.V. 1994. gp74 a membrane glycoprotein of the cis-golgi network that cycles through the endoplasmic reticulum and intermediate compartment. J. Cell Biol. 124 : 649-665 (95) Nobes, C.D., Hall, A. 1995. Rho, Rac and CDC42 GTPases regulate the assembly of multimolecular focal complexes associated with actin stress fibers, lamellipodia and filopodia. Cell 81 : 53-62 (96) Machesky, L., Hall, A. 1996. Rho: a connection between membrane receptor signalling and the cytoskeleton. Trends Cell Biol. 6 : 304-310 (97) Ridley, A.J., Paterson, H.F., Johnston, C.L., Diekmann, D., Hall, A. 1992. The small GTP-binding protein Rac regulates growth factor induced membrane ruffling. Cell 70 : 401-410 (98) Kozma, R., Ahmed, S., Best, A., Lim, L. 1995. The Ras-related protein Cdc42Hs and bradykinin promote formation of peripheral actin microspikes and filopodia in Swiss 3T3 fibroblasts. Mol. Cell Biol. 15 : 1942-1952 (99) Hall, A. 1998. Rho GTPases and the actin cytoskeleton. Science 279 : 509-514 (100) Graf, B., Bähler, M., Hilpelä, P., Böwe, C., Adam, T. 2000. Functional role for the class IX myosin myr5 in epithelial cell infection by Shigella flexneri. Cell. Microbiol. 2 : 601-616 (101) Roux, P., Gauthier-Rouviere, C., Duocet-Brutin, S., Fort, P. 1997. The small GTPases Cdc42Hs, Rac1 and RhoG delineate Raf-independent pathways that cooperate to transform NIH3T3 cells. Curr. Biol. 7 : 629-637 (102) Dumenil, G., Sansonetti, P.J., Tran Van Nhieu, G. 2000. Src tyrosine kinase activity down-regulates rho-dependent responses during Shigella entry into epithelial cells and stress fibre formation. J. Cell Sci. 113 : 71-80 (103) Just, I., Selzer, J., Hofmann, F., Green, G.A., Aktories, K. 1996. Inactivation of Ras by Clostridium sordellii lethal toxin-catalyzed glucosylation. J.Biol.Chem. 271 : 10149-10153 (104) Genth, H., Hofmann, F., Selzer, J., Rex, G., Aktories, K., Just, I. 1996. Difference in protein substrate specificity between hemorrhagic toxin and lethal toxin from Clostridium sordellii . Biochem. Biophys. Res. Commun. 229 : 370-374 (105) Zhang, B., Zheng, Y. 1998. Regulation of RhoA GTP hydrolysis by the GTPase-activating proteins p190, p50RhoGAP, Bcr and 3BP-1. Biochemistry 37 : 5249-5257 (106) Homma, Y., Emori, Y. 1995. A dual functional signal mediator showing RhoGAP and phospholipase C-delta stimulating activities. EMBO J. 14 : 286-291 (107) Chang, J.H., Gill, S., Settleman, J., Parsons, S.J. 1995. C-Src regulates the simultaneous rearrangement of actin cytoskeleton, p190RhoGAP and p120RasGAP following epidermal growth factor stimulation. J. Cell Biol. 130 : 355-368 (108) Fincham, V.J., Chudleigh, A., Frame, M.C. 1999. Regulation of p190 Rho-GAP by v-Src is linked to cytoskeletal disruption during transformation. J. Cell Sci. 112 : 947-956 (109) Dehio, C., Prevost, M.C., Sansonetti, P.J. 1995. Invasion of epithelial cells by Shigella flexneri induces tyrosine phosporylation of cortactin by a pp60c-src-mediated signalling pathway. EMBO J. 14 : 2471-2482 (110) Menkel, A.R., Kroemker, M., Bubeck, P., Ronsiek, M., Nikolai, G., Jockusch, B.M. 1994. Characterization of an F-actin-binding domain in the cytoskeletal protein vinculin. J. Cell Biol. 126 : 1231-1240 (111) Goldmann, W.H., Guttenberg, Z., Tang, J.X., Kroy, K., Isenberg, G., Ezzel, R.M. 1998. Analysis of the F-actin binding fragments of vinculin using stopped-flow and dynamic light-scattering measurements. Eur. J. Biochem. 254 : 413-419 (112) Gilmore, A.P., Burridge, K. 1996. Regulation of vinculin binding to talin and actin by phosphatidyl-inositol-4-5-bisphosphate. Nature 381 : 531-535 (113) Sasaki, T., Takai, Y. 1998. The Rho small G protein family-Rho GDI system as a temporal and spatial determinant for cytoskeletal control. Biochem. Biophys. Res. Commun. 245 : 641-645 (114) Takai, Y., Kaibuchi, K., Sasaki, T., Tanaka, K., Shirataki, H., Nakanishi, H. 1994. Rho small G protein and cytoskeletal control. Princess. Takamatsu. Symp. 24 : 338-350