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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

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