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Temporal and spatial control of nucleophosmin by the Ran–Crm1 complex in centrosome duplication

Nature Cell Biology volume 7, pages 823–830 (2005)Cite this article

Abstract

Centrosome duplication is tightly controlled during faithful cell division, and unnecessary reduplication can lead to supernumerary centrosomes and multipolar spindles that are associated with most human cancer cells1,2,3,4,5. In addition to nucleocytoplasmic transport, the Ran–Crm1 network is involved in regulating centrosome duplication to ensure the formation of a bipolar spindle6,7,8. Here, we discover that nucleophosmin (NPM) may be a Ran–Crm1 substrate that controls centrosome duplication. NPM contains a functional nuclear export signal (NES) that is responsible for both its nucleocytoplasmic shuttling and its association with centrosomes, which are Ran–Crm1-dependent as they are sensitive to Crm1-specific nuclear export inhibition, either by leptomycin B (LMB) or by the expression of a Ran-binding protein, RanBP1. Notably, LMB treatment induces premature centrosome duplication in quiescent cells, which coincides with NPM dissociation from centrosomes. Moreover, deficiency of NPM by RNA interference results in supernumerary centrosomes, which can be reversed by reintroducing wild-type but not NES-mutated NPM. Mutation of a potential proline-dependent kinase phosphorylation site at residue 95, from threonine to aspartic acid (T95D) within the NES motif, abolishes NPM association and inhibition of centrosome duplication. Our results are consistent with the hypothesis that the Ran–Crm1 complex may promote a local enrichment of NPM on centrosomes, thereby preventing centrosome reduplication.

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Figure 1: Nucleocytoplasmic shuttling of NPM depends on the presence of functional NES and NLS motifs mediated by the Ran network.
Figure 2: The centrosome association of NPM and initiation of centrosome duplication are sensitive to Ran–Crm1-specific nuclear export inhibition.
Figure 3: Centrosome-enriched fractions contain NPM, which is sensitive to the nuclear export inhibitor, LMB.
Figure 4: Deficiency of NPM by RNA interference is associated with unscheduled centrosome duplication and abnormal spindles.
Figure 5: The NPM NES motif is required for NPM-mediated suppression of centrosome duplication.

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Acknowledgements

We thank K. Nagata for the generous gift of GFP-tagged NPM, P. Lavia for the RanBP1 expression vector and J. Salisbury for anti-centrin antibody; C. Harris for invaluable comments; S. Garfield, S. Wincovitch and B. J. Taylor for superior technical support; K. MacPherson for bibliographical help; and the National Cancer Institute-Center for Cancer Research Fellows Editorial Board for editorial assistance.

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  1. Liver Carcinogenesis Section, Laboratory of Human Carcinogenesis, Center for Cancer Research, NCI, NIH, Bethesda, 20892, MD, USA

    Wei Wang, Anuradha Budhu, Marshonna Forgues & Xin Wei Wang

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  1. Wei Wang
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  2. Anuradha Budhu
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Correspondence to Xin Wei Wang.

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Wang, W., Budhu, A., Forgues, M. et al. Temporal and spatial control of nucleophosmin by the Ran–Crm1 complex in centrosome duplication. Nat Cell Biol 7, 823–830 (2005). https://doi.org/10.1038/ncb1282

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