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CENP-A octamers do not confer a reduction in nucleosome height by AFM

Nature Structural & Molecular Biology volume 21, pages 4–5 (2014)Cite this article

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Centromeric chromatin comprises nucleosomes with the universal cenH3 (CENP-A) histone variant in place of histone H3. Although there is general agreement as to the function of cenH3 nucleosomes in organizing the kinetochore, the structure of the particle itself has been controversial. One of the methods applied to addressing this issue has been atomic force microscopy (AFM), which measures heights of single nucleosomes on a cleaved mica surface and so can distinguish whether a particle consists of four or eight histones wrapped by DNA. AFM of various reconstituted tetrameric nucleosomes, including cenH3 'hemisomes' (cenH3–H4–H2A–H2B), showed them to be ∼40% shorter than are octameric nucleosomes ('octasomes')1,2, and similar tetrameric dimensions were reported for cenH3 nucleosomes purified from native chromatin3,4,5 (Supplementary Note and Supplementary Fig. 1a).

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Figure 1: Octameric and tetrameric nucleosomes differ in height, but variants do not.

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Acknowledgements

We thank S. Lindsay for helpful discussions and P. Talbert for comments on the manuscript. This work was supported by the Howard Hughes Medical Institute and by a grant from the US National Institutes of Health (U54 CA143862).

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Authors and Affiliations

  1. Basic Sciences Division, Fred Hutchinson Cancer Research Center, Seattle, Washington, USA

    Christine A Codomo, Takehito Furuyama & Steven Henikoff

  2. Howard Hughes Medical Institute, Seattle, Washington, USA

    Takehito Furuyama & Steven Henikoff

Authors
  1. Christine A Codomo
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  2. Takehito Furuyama
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  3. Steven Henikoff
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Correspondence to Steven Henikoff.

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Supplementary information

Supplementary Text and Figures

Supplementary Figure 1 and Supplementary Note (PDF 225 kb)

Supplementary Data Set 1

AFM height measurements (XLSX 111 kb)

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Codomo, C., Furuyama, T. & Henikoff, S. CENP-A octamers do not confer a reduction in nucleosome height by AFM. Nat Struct Mol Biol 21, 4–5 (2014). https://doi.org/10.1038/nsmb.2743

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