Covalent Aurora A regulation by the metabolic integrator coenzyme A

Yugo Tsuchiya,Selena G Burgess,Trang Tran,Alexander Zhyvoloup,Richard Bayliss,Valeriy Filonenko,Yueyang Huang,Alethea B Tabor,Aw Edith Chan,Fiona Bellany,Samantha Ferries,Ivan Gout,Matthias Vonderach,Lalitha Guruprasad,John Carroll,Jovana Baković,Mark Skehel,Patrick A Eyers,Bess Yi Kun Yu,Dominic P Byrne,Sew Y Peak‐Chew ,Jens Bormann ,O M Garifulin

doi:10.1016/j.redox.2019.101318

Abstract

Aurora A kinase is a master mitotic regulator whose functions are controlled by several regulatory interactions and post-translational modifications. It is frequently dysregulated in cancer, making Aurora A inhibition a very attractive antitumor target. However, recently uncovered links between Aurora A, cellular metabolism and redox regulation are not well understood. In this study, we report a novel mechanism of Aurora A regulation in the cellular response to oxidative stress through CoAlation. A combination of biochemical, biophysical, crystallographic and cell biology approaches revealed a new and, to our knowledge, unique mode of Aurora A inhibition by CoA, involving selective binding of the ADP moiety of CoA to the ATP binding pocket and covalent modification of Cys290 in the activation loop by the thiol group of the pantetheine tail. We provide evidence that covalent CoA modification (CoAlation) of Aurora A is specific, and that it can be induced by oxidative stress in human cells. Oxidising agents, such as diamide, hydrogen peroxide and menadione were found to induce Thr 288 phosphorylation and DTT-dependent dimerization of Aurora A. Moreover, microinjection of CoA into fertilized mouse embryos disrupts bipolar spindle formation and the alignment of chromosomes, consistent with Aurora A inhibition.Altogether, our data reveal CoA as a new, rather selective, inhibitor of Aurora A, which locks this kinase in an inactive state via a “dual anchor” mechanism of inhibition that might also operate in cellular response to oxidative stress. Finally and most importantly, we believe that these novel findings provide a new rationale for developing effective and irreversible inhibitors of Aurora A, and perhaps other protein kinases containing appropriately conserved Cys residues.

Highlights

Aurora kinases are Ser/Thr kinases that play well-documented roles in eukaryotes, where they control meiosis, mitosis and cell division [1]
The Aurora kinase domain is itself regulated by a critical autophosphorylation event in the activation loop [8,9,10,11], which is associated with catalytic activation and timely execution of different phases of mitosis [12,13,14,15] and meiosis [16]
We found that Coenzyme A (CoA) is a specific ATP-competitive Aurora A inhibitor in vitro, and employed a combination of biochemical, structural, mass spectrometry and cellular studies to confirm a preferential mode of CoA binding to the active, phosphorylated, Aurora A conformation

Summary

Introduction

Aurora kinases are Ser/Thr kinases that play well-documented roles in eukaryotes, where they control meiosis, mitosis and cell division [1]. The Aurora kinase domain is itself regulated by a critical autophosphorylation event in the activation loop [8,9,10,11], which is associated with catalytic activation and timely execution of different phases of mitosis [12,13,14,15] and meiosis [16]. Aurora B is a chromosomal passenger protein required for the spindle assembly checkpoint and rate-limiting for cytokinesis in cells [20]. Aurora C is most highly expressed in germ cells, where it can functionally replace Auora B as a chromosome passenger protein [1]

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Conclusion