Interaction between Poly(ADP-ribose) and NuMA Contributes to Mitotic Spindle Pole Assembly
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Author(s)
Coughlin, Margaret
Mitchison, Timothy J.
Chang, Paul
Date Issued
September 2009
Journal
Molecular Biology of the Cell
Publisher
American Society for Cell Biology
Citation
Chang, P., M. Coughlin, and T. J. Mitchison. “Interaction between Poly(ADP-ribose) and NuMA Contributes to Mitotic Spindle Pole Assembly.” Molecular Biology of the Cell 20, no. 21 (October 30, 2009): 4575-4585. Copyright © The American Society for Cell Biology
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Final published version
Abstract
Poly(ADP-ribose) (pADPr), made by PARP-5a/tankyrase-1, localizes to the poles of mitotic spindles and is required for bipolar spindle assembly, but its molecular function in the spindle is poorly understood. To investigate this, we localized pADPr at spindle poles by immuno-EM. We then developed a concentrated mitotic lysate system from HeLa cells to probe spindle pole assembly in vitro. Microtubule asters assembled in response to centrosomes and Ran-GTP in this system. Magnetic beads coated with pADPr, extended from PARP-5a, also triggered aster assembly, suggesting a functional role of the pADPr in spindle pole assembly. We found that PARP-5a is much more active in mitosis than interphase. We used mitotic PARP-5a, self-modified with pADPr chains, to capture mitosis-specific pADPr-binding proteins. Candidate binding proteins included the spindle pole protein NuMA previously shown to bind to PARP-5a directly. The rod domain of NuMA, expressed in bacteria, bound directly to pADPr. We propose that pADPr provides a dynamic cross-linking function at spindle poles by extending from covalent modification sites on PARP-5a and NuMA and binding noncovalently to NuMA and that this function helps promote assembly of exactly two poles.
MIT Department
Massachusetts Institute of Technology. Department of Biology
Koch Institute for Integrative Cancer Research at MIT
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DOI of Published Version
https://doi.org/10.1091/mbc.E09-06-0477
