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Polo-like kinase-1 is activated by aurora A to promote checkpoint recovery

Abstract

Polo-like kinase-1 (PLK1) is an essential mitotic kinase regulating multiple aspects of the cell division process1. Activation of PLK1 requires phosphorylation of a conserved threonine residue (Thr 210) in the T-loop of the PLK1 kinase domain, but the kinase responsible for this has not yet been affirmatively identified2,3,4,5,6. Here we show that in human cells PLK1 activation occurs several hours before entry into mitosis, and requires aurora A (AURKA, also known as STK6)-dependent phosphorylation of Thr 210. We find that aurora A can directly phosphorylate PLK1 on Thr 210, and that activity of aurora A towards PLK1 is greatly enhanced by Bora (also known as C13orf34 and FLJ22624), a known cofactor for aurora A (ref. 7). We show that Bora/aurora-A-dependent phosphorylation is a prerequisite for PLK1 to promote mitotic entry after a checkpoint-dependent arrest. Importantly, expression of a PLK1-T210D phospho-mimicking mutant partially overcomes the requirement for aurora A in checkpoint recovery. Taken together, these data demonstrate that the initial activation of PLK1 is a primary function of aurora A.

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Figure 1: Checkpoint recovery requires PLK1 kinase activity.
Figure 2: PLK1 activation occurs several hours before mitosis and requires Thr 210 phosphorylation.
Figure 3: PLK1 activation depends on aurora A.
Figure 4: Aurora-A-dependent phosphorylation of PLK1 is required for recovery.

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Acknowledgements

We are thankful to B. Ingelheim for the gift of BI2536, Millennium Pharmaceuticals for MLN8054, E. Nigg for antibodies and sharing unpublished work, A. Desai and I. Cheeseman for providing the pIC113 plasmid, M. Tanenbaum for U2OS cells expressing YFP–tubulin, S. Sen for GST–aurora A constructs, M. Vromans for recombinant GST–aurora A, Q. Verstralen for technical assistance, D. Littler for active recombinant PLK1, P. de Graaf for the U2OS-TR cell line, G. Kops for making the Deltavision imaging system available, L. Kleij for technical assistance with confocal microscopy, and all members of the Medema laboratory for discussions. We thank M. Alvarez, S. Lens, G. Kops and M. Tanenbaum for critically reading the manuscript. This work was supported by the Dutch Cancer Society (L.M. UU2006-3579), the Dutch Organisation for Scientific Research (R.H.M. ZonMw 918.46.616 and A.L. 916.86.083), the Association for International Cancer Research (L.M. 08-0172) and the NIH (M.B.Y. GM-60594 and CA112967). A.L. was also funded by the Wenner-Gren foundations.

Author Contributions L.M., A.L., D.L., M.A.L., C.C., R.K., R.F. and R.H.M. performed experimental work. All authors were involved in project planning and manuscript preparation.

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Correspondence to René H. Medema.

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The coding sequence for full-length Bora was deposited in the GenBank database (EU834129).

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Macůrek, L., Lindqvist, A., Lim, D. et al. Polo-like kinase-1 is activated by aurora A to promote checkpoint recovery. Nature 455, 119–123 (2008). https://doi.org/10.1038/nature07185

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