Abstract

The widely conserved kinase Aurora B regulates important events during cell division. Surprisingly, recent work has uncovered a few functions of Aurora-family kinases that do not require kinase activity. Thus, understanding this important class of cell cycle regulators will require strategies to distinguish kinase-dependent from independent functions. Here, we address this need in C. elegans by combining germline-specific, auxin-induced Aurora B (AIR-2) degradation with the transgenic expression of kinase-inactive AIR-2. Through this approach, we find that kinase activity is essential for AIR-2’s major meiotic functions and also for mitotic chromosome segregation. Moreover, our analysis revealed insight into the assembly of the ring complex (RC), a structure that is essential for chromosome congression in C. elegans oocytes. AIR-2 localizes to chromosomes and recruits other components to form the RC. However, we found that while kinase-dead AIR-2 could load onto chromosomes, other components were not recruited. This failure in RC assembly appeared to be due to a loss of RC SUMOylation, suggesting that there is crosstalk between SUMOylation and phosphorylation in building the RC and implicating AIR-2 in regulating the SUMO pathway in oocytes. Similar conditional depletion approaches may reveal new insights into other cell cycle regulators.

Highlights

  • The Aurora family of serine/threonine kinases mediates critical cell division events

  • One important protein that is required for both mitotic and meiotic chromosome segregation is the kinase Aurora B, which phosphorylates a variety of other cell division proteins

  • At the restrictive temperature, this mutant exhibited mitotic, but not meiotic defects [9,11,19,20,21]. Consistent with these previous studies, we found that ring complex (RC) and spindle assembly both appear to occur normally in air-2(or207) oocytes at 25 ̊C (S1 Fig)

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Summary

Introduction

The Aurora family of serine/threonine kinases mediates critical cell division events. While Aurora kinases are known to phosphorylate numerous important cell division proteins, a few kinase-independent roles for these proteins have been recently documented. Studies in human cells have shown that the CPC protects centromeric cohesion independent of Aurora B’s kinase activity [2]. In Xenopus egg extracts, the CPC has a kinase-dependent role in phosphorylating outer kinetochore components and a kinase-independent role in ensuring the proper composition of inner kinetochore proteins [3]. Understanding this important family of cell division regulators requires distinguishing between kinase-dependent and independent functions

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