A rho-like protein is involved in the organisation of the contractile ring in dividing sand dollar eggs.
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 cleavage 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 22,000) was ADP-ribosylated by C3 exoenzyme in the isolated cleavage furrow. This protein co-migrated with ADP-ribosylated rhoA derived from human platelets when analysed by two-dimensional gel electrophoresis. These results strongly suggest that a rho-like, small GTP-binding protein is selectively involved in the organisation and maintenance of the contractile ring.
Mabuchi
I
I
Department of Biology, College of Arts and Sciences, University of Tokyo, Japan.
Hamaguchi
Y
Y
Fujimoto
H
H
Morii
N
N
Mishima
M
M
Narumiya
S
S
D036002:ADP Ribose Transferases / Q000494:pharmacology; D000199:Actins / Q000378:metabolism; D000818:Animals; D001905:Botulinum Toxins; D002455:Cell Division / Q000187:drug effects; D005260:Female; D005306:Fertilization / Q000502:physiology; D019204:GTP-Binding Proteins / Q000378:metabolism*; D010063:Ovum / Q000187:drug effects / Q000378:metabolism*; D011065:Poly(ADP-ribose) Polymerases / Q000494:pharmacology; D012617:Sea Urchins / Q000378:metabolism*; D015053:Zygote / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D020741:rho GTP-Binding Proteins
D000199:Actins; D036002:ADP Ribose Transferases; C063007:exoenzyme C3, Clostridium botulinum; D011065:Poly(ADP-ribose) Polymerases; D001905:Botulinum Toxins; D019204:GTP-Binding Proteins; D020741:rho GTP-Binding Proteins