A Meiosis-Specific Form of the APC/C Promotes the Oocyte-to-Embryo Transition by Decreasing Levels of the Polo Kinase Inhibitor Matrimony

Abstract

During the oocyte-to-embryo transition in Drosophila, degradation of the Polo kinase inhibitor, Matrimony, depends on Cortex, a meiosis-specific form of the Anaphase Promoting Complex/Cyclosome that is required for the oocyte's normal transition from meiosis to mitosis.

Abstract

Oocytes are stockpiled with proteins and mRNA that are required to drive the initial mitotic divisions of embryogenesis. But are there proteins specific to meiosis whose levels must be decreased to begin embryogenesis properly? The Drosophila protein Cortex (Cort) is a female, meiosis-specific activator of the Anaphase Promoting Complex/Cyclosome (APC/C), an E3 ubiquitin ligase. We performed immunoprecipitation of Cortex followed by mass spectrometry, and identified the Polo kinase inhibitor Matrimony (Mtrm) as a potential interactor with Cort. In vitro binding assays showed Mtrm and Cort can bind directly. We found Mtrm protein levels to be reduced dramatically during the oocyte-to-embryo transition, and this downregulation did not take place in cort mutant eggs, consistent with Mtrm being a substrate of APC ^Cort. We showed that Mtrm is subject to APC ^Cort-mediated proteasomal degradation and have identified a putative APC/C recognition motif in Mtrm that when mutated partially stabilized the protein in the embryo. Furthermore, overexpression of Mtrm in the early embryo caused aberrant nuclear divisions and developmental defects, and these were enhanced by decreasing levels of active Polo. These data indicate APC ^Cort ubiquitylates Mtrm at the oocyte-to-embryo transition, thus preventing excessive inhibition of Polo kinase activity due to Mtrm's presence.

Author Summary

Despite their many differences, the meiotic and mitotic divisions of the early embryo take place within the same cytoplasmic space. The oocyte-to-embryo transition is the process by which an oocyte, which initially undergoes meiosis, becomes “adapted” to support the rapid mitotic divisions of embryogenesis. This involves fertilization as well as the stockpiling of proteins and mRNA for the transcriptionally silent early embryo. The Anaphase Promoting Complex/Cyclosome (APC/C) is a large protein complex that is active during both mitosis and meiosis and is responsible for targeting certain proteins for degradation. The discovery of the existence of APC/C activators that are present only during meiosis hinted at the possibility that this complex also functions to regulate protein degradation during the oocyte-to-embryo transition. Here we study Cortex, a female- and meiosis-specific activator of the APC/C in the fruit fly Drosophila melanogaster. We find that Cortex activity is necessary for the degradation of Matrimony, a key regulator of female meiosis in Drosophila. Matrimony itself inhibits Polo kinase, another important regulator of both mitosis and meiosis that also functions in chromosome segregation, centrosome dynamics, and cytokinesis. When excess Matrimony protein is not removed from the early embryo, developmental defects arise. Together our findings demonstrate that the precise regulation of Matrimony levels in the egg is necessary for the switch from meiosis to mitosis.