ANALYSIS OF INDUCIBLE CONTRACTILE RINGS SUGGESTS A ROLE FOR PROTEIN-KINASE-C IN EMBRYONIC CYTOKINESIS AND WOUND-HEALING

A semi-in vitro system derived from Xenopus oocytes which allows induction of contractile ring (CR) formation and closure is described and exploited to elucidate regulatory and structural features of cytokinesis. The inducible CRs (ICRs) are composed of actin filaments and closure is actin filament-dependent as is cytokinesis in vivo. ICR closure in this system is calcium-dependent and pH-sensitive, as is cytokinesis in permeabilized cells (Cande: Journal of Cell Biology 87:326, 1980). Closure of ICRs proceeds at a rate and with a kinetic pattern similar to embryonic cytokinesis. Collectively, these data demonstrate that this system is a faithful mimic of cytokinesis in vivo. ICR formation and closure is protein kinase C (PKC)-dependent and neomycin-sensitive, indicating that the PKC branch of the polyphosphoinositide pathway regulates formation of the actomyosin ring which is the effector of cytokinesis. Kinetic measurements show that the rate of ICR closure reaches a peak of 4-8-mu-m/sec. Since the maximum measured velocity of actin filament translocation by vertebrate, non-muscle myosins is 0.04-mu-m/sec, the later observations support a model in which the CR is segmented, containing multiple sites where filaments overlap in a "sliding filament" fashion. Because the rate decreases after reaching a peak, the results also suggest that the number of overlap sites decrease with time.