Deubiquitylation Machinery Is Required for Embryonic Polarity in Caenorhabditis elegans

Abstract

The Caenorhabditis elegans one-cell embryo polarizes in response to a cue from the paternally donated centrosome and asymmetrically segregates cell fate determinants that direct the developmental program of the worm. We have found that genes encoding putative deubiquitylating enzymes (DUBs) are required for polarization of one-cell embryos. Maternal loss of the proteins MATH-33 and USP-47 leads to variable inability to correctly establish and maintain asymmetry as defined by posterior and anterior polarity proteins PAR-2 and PAR-3. The first observable defect is variable positioning of the centrosome with respect to the cell cortex and the male pronucleus. The severity of the polarity defects correlates with distance of the centrosome from the cortex. Furthermore, polarity defects can be bypassed by mutations that bring the centrosome in close proximity to the cortex. In addition we find that polarity and centrosome positioning defects can be suppressed by compromising protein turnover. We propose that the DUB activity of MATH-33 and USP-47 stabilizes one or more proteins required for association of the centrosome with the cortex. Because these DUBs are homologous to two members of a group of DUBs that act in fission yeast polarity, we tested additional members of that family and found that another C. elegans DUB gene, usp-46, also contributes to polarity. Our finding that deubiquitylating enzymes required for polarity in Schizosaccharomyces pombe are also required in C. elegans raises the possibility that these DUBs act through an evolutionarily conserved mechanism to control cell polarity.

Author Summary

In eukaryotes, modification of proteins by the covalent ligation of a protein called ubiquitin is an important regulatory mechanism. In this study we found that deubiquitylation enzymes, which are known to cleave ubiquitin off of target proteins, are required for asymmetry in one-cell embryos of the nematode C. elegans. In one-cell embryos the establishment of asymmetry depends on a signal from the centrosome, a microtubule-organizing center. This signal breaks homogeneity in the contractile cytoskeleton located at the cortex of the embryo. We have identified three deubiquitylation enzymes that are necessary for the centrosome to properly localize adjacent to the cortex to perform its symmetry-breaking role. Furthermore, a homologous group of enzymes in fission yeast also regulates cell polarity. Our results suggest that a novel mechanism of centrosome localization regulated by ubiquitylation exists in C. elegans; this mechanism is masked by genetic redundancy and may be an evolutionarily conserved mechanism for cell asymmetry.