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A rho-like protein is involved in the organisation of the contractile ring in dividing sand dollar eggs

Published online by Cambridge University Press:  26 September 2008

Issei Mabuchi ,
Yukihisa Hamaguchi ,
Hirotaka Fujimoto ,
Narito Morii ,
Masanori Mishima  and
Shuh Narumiya
Issei Mabuchi*
Affiliation:
University of Tokyo, Tokyo Institute of Technology, and Kyoto University, Japan.
Yukihisa Hamaguchi
Affiliation:
University of Tokyo, Tokyo Institute of Technology, and Kyoto University, Japan.
Hirotaka Fujimoto
Affiliation:
University of Tokyo, Tokyo Institute of Technology, and Kyoto University, Japan.
Narito Morii
Affiliation:
University of Tokyo, Tokyo Institute of Technology, and Kyoto University, Japan.
Masanori Mishima
Affiliation:
University of Tokyo, Tokyo Institute of Technology, and Kyoto University, Japan.
Shuh Narumiya
Affiliation:
University of Tokyo, Tokyo Institute of Technology, and Kyoto University, Japan.
*
I. Mabuchi, Department of Biology, College of Arts and Sciences, University of Tokyo, 3-8-1 Komaba, Meguro-Ku, Tokyo, 153, Japan. Tel: 3-3467-1171, ext. 262. Fax: 3-3485-2904.
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Sand dollar eggs were microinjected with botulinum C3 exoenzyme, an ADP-ribosyltransferase from Clostridium botulinum that specifically ADP-ribosylates and inactivates rho proteins. C3 exoenzyme microinjected during nuclear division interfered with subsequent cleavage furrow formation. No actin filaments were detected in the equatorial cortical layer of these eggs by rhodamine-phalloidin staining. When microinjected into furrowing eggs, C3 exoenzyme rapidly disrupted the contractile ring actin filaments and caused regression of the clevage furrows. C3 exoenzyme had no apparent effect on nuclear division, however, and multinucleated embryos developed from the microinjected eggs. By contrast, C3 exoenzyme did not affect the organisation of cortical actin filaments immediately after fertilisation. Only one protein (molecular weight 22000) was ADP-ribosylated by C3 exoenzyme in the isolated cleavage furrow. This protein co-migrated with ADP-ribosylated rhoA derived from human paltelets when analysed by two-dimensional gel electrophoresis. These results strongly suggest that a rho-like, small GTP-binding protein is selectively in the organisation and maintenance of the contractile ring.

Keywords

Actin filamentsC3 exoenzymeCytokinesisSmall G-protein
Type
Article
Information
Zygote , Volume 1 , Issue 4 , November 1993 , pp. 325 - 331
Copyright
Copyright © Cambridge University Press 1993

References

Aktories, K.Weller, U. & Chhatwal, G.S. (1987). Clostridium botulinum type C Produces a novel ADP-ribosyltransferase distinct from botulinum C2 toxin. FEBS Lett. 212, 109–13.CrossRefGoogle ScholarPubMed
Bourne, H.R.Sanders, D.A. & McCormick, F. (1991). The GTPase superfamily: conserved structure and molecular mechanism. Nature 349, 117–27.CrossRefGoogle ScholarPubMed
Chardin, P.Boquet, P.Maduale, P.Popoff, M.R.Rubin, E.J. & Gill, D.M. (1989). The mammalian G protein rhoC is ADP-robosylated by Clostridium botulinum exoenzyme C3 and affects actin microfilaments in Vero cells. EMBO J. 8, 1087–92.CrossRefGoogle Scholar
De Lozanne, A & Spudich, J.A. (1987). Disruption of the Dictyostelium myosin heavy chain gene by homologous recombination. Science 236, 1086–91.CrossRefGoogle ScholarPubMed
Didsbury, J.Weber, R.F.Bokoch, G.M.Evans, T. & Snyderman, R. (1989). rac, a novel ras-related family of proteins that are botulnum toxin substrates. J. Biol. Chem. 264, 16378–82.Google Scholar
Hamaguchi, Y. & Mabuchi, I. (1988). Accumulation of fluorescently labeled actin in the cortical layer in sea urchin egg after fertilization. Cell Motil. Cytoskel. 9, 153–63.CrossRefGoogle ScholarPubMed
Hiramoto, Y. (1974). A method of microinjection. Exp.Cell Res. 87, 403–6.Google Scholar
Kaziro, Y.Ito, H.Kozasa, T.Nakafuku, M. & Sato, T. (1991) Structure and function of signal-transducing GTP-binding Proteins. Annu. Rev. Biochem. 60, 349–400.CrossRefGoogle ScholarPubMed
Kishi, K.Sasaki, T.Kuroda, S.Itoh, T. & Takai, Y. (1993). Regulation of cytoplasmic division of Xenopus embryo by rho p21 and its inhibitory GDP/GTP exchange protein (rho GDI). J. Cell Biol. 120, 1187–95.Google Scholar
Knecht, D.A. & Loomis, W.F. (1987). Antisense RNA inactivation of myosin heavy chain gene expression in Dictyostelium discoideum. Science 236, 1081–6.Google Scholar
Mabuchi, I. (1986). Biochmical aspects of cytokinesis. Int. Rev. Cytol. 101, 175–213.CrossRefGoogle Scholar
Mabuchi, I. & Okuno, M. (1977). The effect of myosin antibody on the division of starfish blastomeres. J. Cell Biol. 74, 251–63.CrossRefGoogle ScholarPubMed
Mabuchi, I. & Takano-Ohmuro, H. (1990). Effects of inhibitors of myosin light chain kinase and other protein kinases on the first cell division of sea urchin eggs. Dev. Growth Differ. 32, 549–56.CrossRefGoogle ScholarPubMed
Mabuchi, I.Tsukita, S.Tsukita, S. & Sawai, T. (1988). Cleavage furrow isolated from newt eggs: contraction, organization of actin filaments, and protein components of the furrow. Proc. Natl. Acad. Sci. USA 86, 5966–70.Google Scholar
Menard, L.Tomhave, E.Casey, P.J.Uhing, R.J.Snyderman, R. & Didsbury, J.R. (1992). Racl, a low-molecular-mass, GTP-binding-protein with high intrinsic GTPase activity and distinct biochemical properties. Eur. J. Biochem. 206, 537–46.Google Scholar
Morii, N.Sekine, A.Ohashi, Y.Nakao, K.Imura, H.Fujiwara, M. & Narumiya, S. (1988). Purification and properties of the cytosolic substrate for botulinum ADP-ribosultransferase. J. Biol. Chem. 263, 12420–6.Google Scholar
Morii, N.Kawano, K.Sekine, A.Yamada, T. & Narumiya, S. (1991). purification of GTPase-activating protein specific for the rho gene products. J. Biol. Chem. 266, 7646–50.Google Scholar
Morii, N.Teru-uchi, T.Tominaga, T.Kumagai, N.Kozaki, S.Ushikubi, F. & Narumiya, S. (1992). A rho gene product in human blood platelets. J. Biol. Chem. 267, 20921–6.CrossRefGoogle ScholarPubMed
Narumiya, S. & Morii, N. (1993). rho gene products, botulinum C3 exoenzyme and cell adhesion. Cellular Signalling 5, 9–19.CrossRefGoogle ScholarPubMed
Nemoto, Y.Namba, T.Kozaki, S. & Narumiya, S. (1991). Clostridium botulinum C3 ADP-ribosyltransferase gene. Cloning, sequencing, and expression of a functional protein in Escherichia coli. J. Biol. Chem. 266, 19312–19.CrossRefGoogle ScholarPubMed
Nemoto, Y.Namba, T.Teru-uchi, T.Ushikubi, F.Morii, N. & Narumiya, S. (1992). A rho gene product in human blood platelets. I. Identification of the platelet substrate for botulinum C3 ADP-ribosyltransferase as rhoA protein. J. Biol. Chem. 267, 20916–20.Google Scholar
O'Farrell, P.Z.Goodman, H.M. & O'Farrell, P.H. (1977). High resolution two-dimensional electrophoresis of basic as well as acidic proteins. Cell 12, 1133–42.CrossRefGoogle ScholarPubMed
Paterson, H.F.Self, A.L.Garrett, M.D.Just, I.Aktories, K. & Hall, A. (1990). Microinjection of recombinant p21rho induces rapid changes in cell morphology. J. Cell Biol. 111, 1001–7.Google Scholar
Rappaport, R. (1986). Establishment of the mechanism of cytokinesis in animal cells. Int. Rev. Cytol. 105, 245–81.CrossRefGoogle ScholarPubMed
Ridley, A.J. & Hall, A. (1992). The small GTP-binding protein rho regulates the assembly of focal adhesions and actin stress fibers in response to growth factors. Cell 70, 389–99.Google Scholar
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, 401–10.CrossRefGoogle ScholarPubMed
Rubin, E.L.Gill, D.M.Boquet, P. & Popoff, M.R. (1988). Functional modification of a 21-Kilodalton G protein when ADP-ribosylated by exoenzyme C3 of Clostridium botulinum. Mol. Cell Biol. 8, 418–26.Google Scholar
Schroeder, T.F. (1972). The Contractile ring. II. Determiniing its brief existence, volumetric changes, and vital role in cleaving Arbacia eggs. J. Cell Biol. 53, 419–34.Google Scholar
Sekine, A.Fujiwara, M. & Narumiya, S. (1989). Asparagine residue in the rho gene product is the modification site for botulinum ADP-ribosyltransferase. J. Biol. Chem. 264, 8602–5.CrossRefGoogle ScholarPubMed
Tominaga, T.Sugie, K.Hirata, M.Morii, N.Fukada, J.Uchida, A.Imura, H. & Narumiya, S. (1993). Inhibition of PMA-induced, LFA-1 dependent lymphocyte aggregation by ADP ribosylation of the small molecular weight GTP binding protein,. rho.J. Cell Biol. 120, 1529–37.CrossRefGoogle ScholarPubMed
Yonemura, S. & Mabuchi, I. (1987). Wave of cortical actin polymerization in the sea urchin egg. Cell Motil. Cytoskel.. 7, 46–53.Google Scholar
Yonemura, S.Mabuchi, I. & Tsukita, S. (1991). Mass isolation of cleavege furrows from dividing sea urchin eggs. J. Cell Sci.. 100, 73–84.Google Scholar