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Tankyrase-1 polymerization of poly(ADP-ribose) is required for spindle structure and function

Nature Cell Biology volume 7, pages 1133–1139 (2005)Cite this article

Abstract

Poly(ADP-ribose) (PAR) is a large, negatively charged post-translational modification that is produced by polymerization of NAD+ by PAR polymerases (PARPs)1. There are at least 18 PARPs in the human genome, several of which have functions that are unknown1. PAR modifications are dynamic; PAR structure depends on the balance between synthesis and hydrolysis by PAR glycohydrolase2. We previously found that PAR is enriched in vertebrate somatic-cell mitotic spindles and demonstrated a requirement for PAR in the assembly of Xenopus egg extract spindles3. Here, we knockdown all characterized PARPs using RNA interference (RNAi), and identify tankyrase-1 as the PARP that is required for mitosis. Tankyrase-1 localizes to mitotic spindle poles, to telomeres4 and to the Golgi apparatus5. Tankyrase-1 RNAi was recently shown to result in mitotic arrest, with abnormal chromosome distributions and spindle morphology observed — data that is interpreted as evidence of post-anaphase arrest induced by failure of telomere separation6. We show that tankyrase-1 RNAi results in pre-anaphase arrest, with intact sister-chromatid cohesion. We also demonstrate a requirement for tankyrase-1 in the assembly of bipolar spindles, and identify the spindle-pole protein NuMA7 as a substrate for covalent modification by tankyrase-1.

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Figure 1: Tankyrase-1 PARP activity is required for spindle structure and function.
Figure 2: Tankyrase-1 knockdown activates the Mad2-dependent spindle checkpoint and results in pre-anaphase mitotic arrest.
Figure 3: Tankyrase-1 knockdown results in centrosome-independent spindle-pole assembly.
Figure 4: Tankyrase-1 is required for spindle-pole function and spindle bipolarity and structure.
Figure 5: The spindle-pole protein NuMA is PARsylated by tankyrase-1 in vivo.

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Acknowledgements

We thank Z. Perlman for critical evaluation of the manuscript. We thank A. Groen, P. Ohi and V. Kickhoefer for antibodies, and D. Compton for antibodies and DNA. We thank T. De Lange for tankyrase-1 clones. This work was supported by a National Institutes of Health grant (GM39565) to T.J.M. and a National Institute of Health Postdoctoral fellowship to P.C.

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  1. Department of Systems Biology, Harvard Medical School, Boston, 02115, MA, USA

    Paul Chang, Margaret Coughlin & Timothy J. Mitchison

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Correspondence to Paul Chang.

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Chang, P., Coughlin, M. & Mitchison, T. Tankyrase-1 polymerization of poly(ADP-ribose) is required for spindle structure and function. Nat Cell Biol 7, 1133–1139 (2005). https://doi.org/10.1038/ncb1322

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