Caenorhabditis elegans embryonic polarity requires the asymmetrically distributed proteins PAR-3, PAR-6 and PKC-3. The rho family GTPase CDC-42 regulates the activities of these proteins in mammals, flies and worms. To clarify its mode of action in C. elegans we disrupted the interaction between PAR-6 and CDC-42 in vivo, and also determined the distribution of GFP-tagged CDC-42 in the early embryo. Mutant PAR-6 proteins unable to interact with CDC-42 accumulated asymmetrically, at a reduced level, but this asymmetry was not maintained during the first division. We also determined that constitutively active GFP∷CDC-42 becomes enriched in the anterior during the first cell cycle in a domain that overlaps with PAR-6. The asymmetry is dependent on PAR-2, PAR-5 and PAR-6. Furthermore, we found that overexpression of constitutively active GFP∷CDC-42 increased the size of the anterior domain. We conclude that the CDC-42 interaction with PAR-6 is not required for the initial establishment of asymmetry but is required for maximal cortical accumulation of PAR-6 and to maintain its asymmetry.
