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CENP-A confers a reduction in height on octameric nucleosomes

Abstract

Nucleosomes with histone H3 replaced by CENP-A direct kinetochore assembly. CENP-A nucleosomes from human and Drosophila have been reported to have reduced heights as compared to canonical octameric H3 nucleosomes, thus suggesting a unique tetrameric hemisomal composition. We demonstrate that octameric CENP-A nucleosomes assembled in vitro exhibit reduced heights, indicating that they are physically distinct from H3 nucleosomes and negating the need to invoke the presence of hemisomes.

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Figure 1: In vitro–assembled histone H3 and CENP-A nucleosomes behave as octamers.
Figure 2: Octameric CENP-A nucleosomes are lower in height.

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Acknowledgements

The authors acknowledge A. Kerr for performing statistical analysis on AFM data and M. Barrios-Llerena for performing MS analyses. The authors thank B.E. Black (University of Pennsylvania, Philadelphia, Pennsylvania, USA) for sharing results prior to publication. AFM was enabled by the Bioimaging Small Research Facility, School of Engineering, University of Edinburgh. The Wellcome Trust supported the work of R.C.A. (095021 and 065061), J.R. (084229) and T.O.-H. (095062) and the Edinburgh Protein Production Facility (081287). Work in the Wellcome Trust Centre for Cell Biology is supported by the Wellcome Trust core funding (092076). M.D.D.M. was supported by a Scottish University Life Sciences Alliance Prize studentship. A.F.S. was supported by the US National Institutes of Health (R01 GM074728), C.J.F. by the Stanford Lieberman Fellowship and A.G. by the German Research Foundation. R.C.A. was supported as a Wellcome Trust Principal Fellow.

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Contributions

M.D.D.M. jointly conceived of the study with R.C.A.; M.D.D.M. produced the S. pombe nucleosome arrays with assistance from T.O.-H. and performed the AFM experiments with assistance from A.D.; C.J.F. and A.G. produced the human nucleosome arrays with assistance from A.F.S.; A.G. and M.D.D.M. jointly performed AvaI digestion experiments; C.J.F. and M.D.D.M. jointly performed MNase digestion experiments; and H.M.B. and M.D.D.M. jointly performed the fixation experiments. J.R. assisted with performing the MS analysis. M.D.D.M. prepared the manuscript, which was edited by A.F.S. and R.C.A. All authors discussed the results and implications and commented on the manuscript at all stages.

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Correspondence to Robin C Allshire.

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The authors declare no competing financial interests.

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Miell, M., Fuller, C., Guse, A. et al. CENP-A confers a reduction in height on octameric nucleosomes. Nat Struct Mol Biol 20, 763–765 (2013). https://doi.org/10.1038/nsmb.2574

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