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Nucleic Acids Research Advance Access originally published online on March 31, 2009
Nucleic Acids Research 2009 37(10):3391-3406; doi:10.1093/nar/gkp199
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Nucleic Acids Research, 2009, Vol. 37, No. 10 3391-3406
© 2009
The Author(s) This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Gene Regulation, Chromatin and Epigenetics

Linker histone H1 is present in centromeric chromatin of living human cells next to inner kinetochore proteins

S. Orthaus1, K. Klement1, N. Happel2, C. Hoischen1 and S. Diekmann1,*

1Leibniz-Institute for Age Research - Fritz Lipmann Institute, Beutenbergstr. 11, D-07745 Jena and 2Department of Molecular Biology, Institute for Biochemistry and Molecular Cell Biology, University Goettingen, Humboldtallee 23, D-37073 Goettingen, Germany

*To whom correspondence should be addressed. Tel: +49 3641 65 6260; Fax: +49 3641 65 6261; Email: diekmann{at}fli-leibniz.de

Received November 11, 2008. Revised March 9, 2009. Accepted March 10, 2009.

The vertebrate kinetochore complex assembles at the centromere on {alpha}-satellite DNA. In humans, {alpha}-satellite DNA has a repeat length of 171 bp slightly longer than the DNA in the chromatosome containing the linker histone H1. The centromere-binding protein CENP-B binds specifically to {alpha}-satellite DNA with properties of a centromeric-linker histone. Here, we analysed if linker histone H1 is present at or excluded from centromeric chromatin by CENP-B. By immunostaining we detected the presence, but no enrichment or depletion of five different H1 subtypes at centromeric chromatin. The binding dynamics of H1 at centromeric sites were similar to that at other locations in the genome. These dynamics did not change in CENP-B depleted cells, suggesting that CENP-B and H1 co-exist in centromeric chromatin with no or little functional overlap. By bimolecular fluorescence complementation (BiFC) and Förster resonance energy transfer (FRET), we revealed that the linker histone H1 subtypes H1° and H1.2 bind to centromeric chromatin in interphase nuclei in direct neighbourhood to inner kinetochore proteins.


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