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Silencing of Nuclear Mitotic Apparatus protein (NuMA) accelerates the apoptotic disintegration of the nucleus

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Abstract

One main feature of apoptosis is the sequential degradation of the nuclear structure, including the fragmentation of chromatin and caspase-mediated cleavage of various nuclear proteins. Among these proteins is the Nuclear Mitotic Apparatus protein (NuMA) which plays a specific role in the organization of the mitotic spindle. The exact function of NuMA in the interphase nucleus is unknown, but a number of reports have suggested that it may play a role in chromatin organization and/or gene expression. Here we show that upon cleavage in apoptotic cells, the N-terminal cleavage fragment of NuMA is solubilized while the C-terminal fragment remains associated with the condensed chromatin. Using pancaspase inhibitor z-VAD-fmk and caspase-3 deficient MCF-7 cells, we further show that the solubilization is dependent on caspase-mediated cleavage of NuMA. Finally, the silencing of NuMA by RNAi accelerated nuclear breakdown in apoptotic MCF-7 cells. These results suggest that NuMA may provide structural support in the interphase nucleus by contributing to the organization of chromatin.

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Acknowledgements

We would like to thank Professor D. Compton for donating the GFP constructs and Dr. A. Merdes for providing the sequence for NuMA siRNA oligonucleotide. Dr. Gudrun Wahlström is thanked for her help and expertise in the GFP work. Harry Kujari and Jaakko Matomäki are thanked for their help in the statistical analysis of the data. Dr. Ellen Valle is thanked for her help in the proof-reading of the text. This work was supported by the Finnish Medical Foundation, the Finnish-Norwegian Medical Foundation, the Emil Aaltonen Foundation, the Emil and Blida Maunula Trust of the University of Turku, the EVO Grant of the Turku University Central Hospital, the Turku University Foundation, Valto Takala Trust, The Varsinais-Suomi Regional Fund of the Finnish Cultural Foundation, the Medical Faculty of the University of Turku and the Turku Graduate School of Biomedical Sciences (TuBS).

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The authors declare no conflict of interest.

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Authors and Affiliations

  1. Department of Pathology, University of Turku, Kiinamyllynkatu 10, FI-20520, Turku, Finland

    Katri Kivinen, Pekka Taimen & Markku Kallajoki

  2. Medicity Research Laboratory, University of Turku, Tykistökatu 6 A, FI-20520, Turku, Finland

    Katri Kivinen

  3. Turku Graduate School of Biomedical Sciences, Turku, Finland

    Katri Kivinen

  4. Department of Pathology, Turku University Hospital, FI-20520, Turku, Finland

    Pekka Taimen & Markku Kallajoki

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  1. Katri Kivinen
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Correspondence to Katri Kivinen.

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Kivinen, K., Taimen, P. & Kallajoki, M. Silencing of Nuclear Mitotic Apparatus protein (NuMA) accelerates the apoptotic disintegration of the nucleus. Apoptosis 15, 936–945 (2010). https://doi.org/10.1007/s10495-010-0506-8

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