Covalent Aurora A regulation by the metabolic integrator coenzyme A
Yugo Tsuchiya,
Dominic P. Byrne,
Selena G. Burgess,
Jenny Bormann,
Jovana Baković,
Yueyang Huang,
Alexander Zhyvoloup,
Bess Yi Kun Yu,
Sew Peak-Chew,
Trang Tran,
Fiona Bellany,
Alethea B. Tabor,
AW Edith Chan,
Lalitha Guruprasad,
Oleg Garifulin,
Valeriy Filonenko,
Matthias Vonderach,
Samantha Ferries,
Claire E. Eyers,
John Carroll,
Mark Skehel,
Richard Bayliss,
Patrick A. Eyers,
Ivan Gout
Affiliations
Yugo Tsuchiya
Department of Structural and Molecular Biology, University College London, London, WC1E 6BT, UK
Dominic P. Byrne
Department of Biochemistry, Institute of Integrative Biology, University of Liverpool, Liverpool, L69 7ZB, UK
Selena G. Burgess
School of Molecular and Cellular Biology, Astbury Centre for Structural and Molecular Biology, University of Leeds, Leeds, LS2 9JT, UK
Jenny Bormann
Department of Cell and Developmental Biology, University College London, London, WC1E 6BT, UK
Jovana Baković
Department of Structural and Molecular Biology, University College London, London, WC1E 6BT, UK
Yueyang Huang
Department of Structural and Molecular Biology, University College London, London, WC1E 6BT, UK
Alexander Zhyvoloup
Department of Structural and Molecular Biology, University College London, London, WC1E 6BT, UK
Bess Yi Kun Yu
Department of Structural and Molecular Biology, University College London, London, WC1E 6BT, UK
Sew Peak-Chew
MRC Laboratory of Molecular Biology, Cambridge Biomedical Campus, Cambridge, CB2 0QH, UK
Trang Tran
Department of Chemistry, University College London, London, WC1E 6BT, UK
Fiona Bellany
Department of Chemistry, University College London, London, WC1E 6BT, UK
Alethea B. Tabor
Department of Chemistry, University College London, London, WC1E 6BT, UK
AW Edith Chan
Wolfson Institute for Biomedical Research, University College London, London, WC1E 6BT, UK
Lalitha Guruprasad
School of Chemistry, University of Hyderabad, Hyderabad, 500 046, India
Oleg Garifulin
Department of Cell Signaling, Institute of Molecular Biology and Genetics, Kyiv 143, Ukraine
Valeriy Filonenko
Department of Cell Signaling, Institute of Molecular Biology and Genetics, Kyiv 143, Ukraine
Matthias Vonderach
Centre for Proteome Research, Department of Biochemistry, Institute of Integrative Biology, University of Liverpool, Liverpool, L69 7ZB, UK
Samantha Ferries
Centre for Proteome Research, Department of Biochemistry, Institute of Integrative Biology, University of Liverpool, Liverpool, L69 7ZB, UK
Claire E. Eyers
Department of Biochemistry, Institute of Integrative Biology, University of Liverpool, Liverpool, L69 7ZB, UK; Centre for Proteome Research, Department of Biochemistry, Institute of Integrative Biology, University of Liverpool, Liverpool, L69 7ZB, UK
John Carroll
Department of Cell and Developmental Biology, University College London, London, WC1E 6BT, UK
Mark Skehel
MRC Laboratory of Molecular Biology, Cambridge Biomedical Campus, Cambridge, CB2 0QH, UK
Richard Bayliss
School of Molecular and Cellular Biology, Astbury Centre for Structural and Molecular Biology, University of Leeds, Leeds, LS2 9JT, UK; Corresponding author.
Patrick A. Eyers
Department of Biochemistry, Institute of Integrative Biology, University of Liverpool, Liverpool, L69 7ZB, UK; Corresponding author.
Ivan Gout
Department of Structural and Molecular Biology, University College London, London, WC1E 6BT, UK; Department of Cell Signaling, Institute of Molecular Biology and Genetics, Kyiv 143, Ukraine; Corresponding author. Department of Structural and Molecular Biology, University College London, London, WC1E 6BT, UK.
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.
