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Polo-like kinase 1 phosphorylates cyclin B1 and targets it to the nucleus during prophase

An Erratum to this article was published on 12 April 2001

Abstract

In vertebrate cells, the nuclear entry of Cdc2–cyclin B1 (MPF1,2,3) during prophase4,5,6 is thought to be essential for the induction and coordination of M-phase events7,8,9,10,11,12. Phosphorylation of cyclin B1 is central to its nuclear translocation8,13,14, but the kinases that are responsible remain unknown. Here we have purified a protein kinase from Xenopus M-phase extracts that phosphorylates a crucial serine residue (S147) in the middle of the nuclear export signal sequence10,13,15 of cyclin B1. We have identified this kinase as Plx1 (ref. 16), a Xenopus homologue of Polo-like kinase (Plk)-1 (refs 17, 18). During cell-cycle progression in HeLa cells, a change in the kinase activity of endogenous Plk1 toward S147 and/or S133 correlates with a kinase activity in the cell extracts. An anti-Plk1 antibody depletes the M-phase extracts of the kinase activity toward S147 and/or S133. An anti-phospho-S147 antibody reacts specifically with cyclin B1 only during G2/M phase. A mutant cyclin B1 in which S133 and S147 are replaced by alanines remains in the cytoplasm, whereas wild-type cyclin B1 accumulates in the nucleus during prophase. Co-expression of constitutively active Plk1 stimulates nuclear entry of cyclin B1. Our results indicate that Plk1 may be involved in targeting MPF to the nucleus during prophase.

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Figure 1: Phosphorylation-dependent nuclear localization of cyclin B1.
Figure 2: A kinase activity that phosphorylates S133 and/or S147 of cyclin B1 is activated during the G2/M transition.
Figure 3: Purification of a kinase activity that phosphorylates S133 and/or S147 of cyclin B1.
Figure 4: Plk1 phosphorylates cyclin B1 on S133 and/or S147 during G2/M phase in HeLa cells.
Figure 5: Plk1-mediated phosphorylation of cyclin B1 on S147 in vitro and in vivo.
Figure 6: Constitutively active Plk1 induces nuclear localization of cyclin B1 in the absence of Cdc2 kinase activity.

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Acknowledgements

We are grateful to K. Todokoro for a mouse Plk1 complementary DNA and to H. Ellinger-Ziegelbauer for advice on Plk1 assays. We thank M. Yanagida for helpful discussion. This work was supported by grants from the Ministry of Education, Science and Culture of Japan (E.N.).

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Correspondence to Eisuke Nishida.

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Toyoshima-Morimoto, F., Taniguchi, E., Shinya, N. et al. Polo-like kinase 1 phosphorylates cyclin B1 and targets it to the nucleus during prophase. Nature 410, 215–220 (2001). https://doi.org/10.1038/35065617

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