Abstract
The centromere is the chromosomal locus that ensures fidelity in genome transmission at cell division. Centromere protein A (CENP-A) is a histone H3 variant that specifies centromere location independently of DNA sequence. Conflicting evidence has emerged regarding the histone composition and stoichiometry of CENP-A nucleosomes. Here we show that the predominant form of the CENP-A particle at human centromeres is an octameric nucleosome. CENP-A nucleosomes are very highly phased on α-satellite 171-base-pair monomers at normal centromeres and also display strong positioning at neocentromeres. At either type of functional centromere, CENP-A nucleosomes exhibit similar DNA-wrapping behavior, as do octameric CENP-A nucleosomes reconstituted with recombinant components, having looser DNA termini than those on conventional nucleosomes containing canonical histone H3. Thus, the fundamental unit of the chromatin that epigenetically specifies centromere location in mammals is an octameric nucleosome with loose termini.
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Acknowledgements
We thank O. Jabado (Icahn School of Medicine at Mount Sinai, New York, New York, USA) for help with Illumina sequencing, T. Patel (University of Pennsylvania, Philadelphia, Pennsylvania, USA (UPenn)) and B. Cole (UPenn) for advice on data analysis, D. Rogers (UPenn) and B. Gregory (UPenn) for advice, E. Bernstein (Icahn School of Medicine at Mount Sinai, New York, New York, USA) for mentoring and advising D.H., M. Lampson (UPenn) for comments on the manuscript, K. Luger (Colorado State University, Fort Collins, Colorado, USA), D. Cleveland (University of California, San Diego, La Jolla, California, USA), A. Straight (Stanford University, Stanford, California, USA) and D. Rhodes (Medical Research Council Laboratory of Molecular Biology, Cambridge, UK) for plasmids, A. Choo (Murdoch Children's Research Institute, Victoria, Australia) for the cell line containing the PD-NC4 chromosome and R. Allshire (University of Edinburgh, Edinburgh, Scotland, UK) for sharing results before publication. This work was supported by US National Institutes of Health research grant GM082989 (B.E.B.), a Career Award in the Biomedical Sciences from the Burroughs Wellcome Fund (B.E.B.), a Rita Allen Foundation Scholar Award (B.E.B.), a predoctoral fellowship from the American Heart Association (K.J.S.) and a postdoctoral fellowship from the American Cancer Society (N.S.). T.P. acknowledges support from US National Institutes of Health grant GM08275 (UPenn Structural Biology Training Grant).
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D.H. designed and performed experiments and analyzed data. K.J.S. and T.P. designed and performed experiments, developed new analytical tools, analyzed data and wrote the manuscript. M.U.S. developed new analytical tools. N.S. analyzed data and modeled nucleosomes. A.A. performed experiments and provided technical advice on ChIP experiments. P.E.W. directed the project, designed experiments and analyzed data. B.E.B. directed the project, designed experiments, analyzed data and wrote the manuscript.
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Hasson, D., Panchenko, T., Salimian, K. et al. The octamer is the major form of CENP-A nucleosomes at human centromeres. Nat Struct Mol Biol 20, 687–695 (2013). https://doi.org/10.1038/nsmb.2562
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DOI: https://doi.org/10.1038/nsmb.2562
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