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Formation of stable attachments between kinetochores and microtubules depends on the B56-PP2A phosphatase

Nature Cell Biology volume 13, pages 1265–1271 (2011)Cite this article

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Abstract

Error-free chromosome segregation depends on the precise regulation of phosphorylation to stabilize kinetochore–microtubule attachments (K-fibres) on sister chromatids that have attached to opposite spindle poles (bi-oriented)1. In many instances, phosphorylation correlates with K-fibre destabilization2,3,4,5,6,7. Consistent with this, multiple kinases, including Aurora B and Plk1, are enriched at kinetochores of mal-oriented chromosomes when compared with bi-oriented chromosomes, which have stable attachments2,8. Paradoxically, however, these kinases also target to prometaphase chromosomes that have not yet established spindle attachments and it is therefore unclear how kinetochore–microtubule interactions can be stabilized when kinase levels are high. Here we show that the generation of stable K-fibres depends on the B56-PP2A phosphatase, which is enriched at centromeres/kinetochores of unattached chromosomes. When B56-PP2A is depleted, K-fibres are destabilized and chromosomes fail to align at the spindle equator. Strikingly, B56-PP2A depletion increases the level of phosphorylation of Aurora B and Plk1 kinetochore substrates as well as Plk1 recruitment to kinetochores. Consistent with increased substrate phosphorylation, we find that chemical inhibition of Aurora or Plk1 restores K-fibres in B56-PP2A-depleted cells. Our findings reveal that PP2A, an essential tumour suppressor9, tunes the balance of phosphorylation to promote chromosome–spindle interactions during cell division.

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Figure 1: Microtubule-sensitive targeting of PP2A to centromeres/kinetochores during cell division.
Figure 2: Microtubule-sensitive targeting of B56 regulatory subunits to centromeres/kinetochores.
Figure 3: B56-PP2A is required for stable kinetochore–microtubule attachments and chromosome alignment.
Figure 4: B56-PP2A depletion increases the level of phosphorylation of Aurora B substrates and Aurora inhibition suppresses the B56-PP2A-siRNA phenotype.
Figure 5: B56-PP2A regulates Plk1 substrate phosphorylation and Plk1 targeting to the kinetochore, and Plk1 inhibition suppresses the B56-PP2A-siRNA phenotype.

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Acknowledgements

We thank I. Cheeseman (MIT, Cambridge, Massachusetts, USA), A. Desai (University of California San Diego, San Diego, California, USA), S. Elowe (Centre de Recherche du CHUQ, Quebec, Canada) and H. Yu (UT Southwestern Medical Center, Dallas, Texas, USA) for antibodies and W. Harper for plasmids. E.A.F. was supported by a Damon Runyon Cancer Research postdoctoral fellowship (DRG 1936-07) and the Charles H. Revson Foundation. This work was supported by NIH GM65933 (T.M.K.).

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  1. Laboratory of Chemistry and Cell Biology, Rockefeller University, 1230 York Avenue, New York, New York 10065, USA

    Emily A. Foley, Maria Maldonado & Tarun M. Kapoor

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E.A.F. and T.M.K. designed the experiments and wrote the paper. E.A.F. carried out essentially all of the experiments. M.M. contributed to the live-cell imaging.

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Correspondence to Tarun M. Kapoor.

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Foley, E., Maldonado, M. & Kapoor, T. Formation of stable attachments between kinetochores and microtubules depends on the B56-PP2A phosphatase. Nat Cell Biol 13, 1265–1271 (2011). https://doi.org/10.1038/ncb2327

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