Abstract
The considerable length of DNA in eukaryotic genomes requires packaging into chromatin to fit inside the small dimensions of the cell nucleus. Histone H1 functions in the compaction of chromatin into higher order structures derived from the repeating ‘beads on a string’ nucleosome polymer. Modulation of H1 binding activity is thought to be an important step in the potentiation/depotentiation of chromatin structure for transcription1,2,3,4. It is generally accepted that H1 binds less tightly than other histones to DNA in chromatin and can readily exchange in living cells5,6,7,8. Fusion proteins of Histone H1 and green fluorescent protein (GFP) have been shown9 to associate with chromatin in an apparently identical fashion to native histone H1. This provides a means by which to study histone H1–chromatin interactions in living cells. Here we have used human cells with a stably integrated H1.1–GFP fusion protein to monitor histone H1 movement directly by fluorescence recovery after photobleaching in living cells. We find that exchange is rapid in both condensed and decondensed chromatin, occurs throughout the cell cycle, and does not require fibre–fibre interactions. Treatment with drugs that alter protein phosphorylation significantly reduces exchange rates. Our results show that histone H1 exchange in vivo is rapid, occurs through a soluble intermediate, and is modulated by the phosphorylation of a protein or proteins as yet to be determined.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Zlatanova, J. & Van Holde, K. Histone H1 and transcription: still an enigma? J. Cell Sci. 103, 889– 895 (1992).
Shen, X. & Gorovsky, M. A. Linker histone H1 regulates specific gene expression but not global transcription in vivo. Cell 86, 475–483 ( 1996).
Wolffe, A. P., Khochbin, S. & Dimitrov, S. What do linker histones do in chromatin? BioEssays 19, 249–255 ( 1997).
Thomas, J. O. Histone H1: location and role. Curr. Opin. Cell Biol. 11, 312–317 (1999).
Thomas, J. O. & Rees, C. Exchange of histones H1 and H5 between chromatin fragments. A preference of H5 for higher-order structures. Eur. J. Biochem. 134, 109–115 (1983).
Louters, L. & Chalkley, R. Exchange of histones H1, H2A, and H2B in vivo. Biochemistry 24, 3080 –3085 (1985).
Wu, L. H., Kuehl, L. & Rechsteiner, M. Dynamic behavior of histone H1 microinjected into HeLa cells. J. Cell Biol. 103, 465– 474 (1986).
Hendzel, M. J. & Davie, J. R. Nucleosomal histones of transcriptionally active/competent chromatin preferentially exchange with newly synthesized histones in quiescent chicken erythrocytes. Biochem. J. 271, 67–73 (1990).
Gunjan, A., Alexander, B. T., Sittman, D. B. & Brown, D. T. Effects of H1 histone variant overexpression on chromatin structure. J. Biol. Chem. 274, 37950–37956 (1999).
Kruhlak, M. Reduced mobility of the ASF splicing factor in the nucleoplasm and through steady-state speckle compartments. J. Cell Biol. 150 , 41–51 (2000).
Li, W., Nagaraja, S., Delcuve, G. P., Hendzel, M. J. & Davie, J. R. Effects of histone acetylation, ubiquitination and variants on nucleosome stability. Biochem. J. 296, 737–744 ( 1993).
Phair, R. D. & Misteli, T. High mobility of proteins in the mammalian cell nucleus. Nature 404, 604– 609 (2000).
Jin, Y. J. & Cole, R. D. Exchange of H1 histone depends on aggregation of chromatin, not simply on ionic strength. J. Biol. Chem. 261, 15805–15812 ( 1986).
Lu, M. J. et al. Generation and characterization of novel antibodies highly selective for phosphorylated linker histone H1 in Tetrahymena and HeLa cells. Chromosoma 103, 111–121 (1994).
Th’ng, J. P., Guo, X. W., Swank, R. A., Crissman, H. A. & Bradbury, E. M. Inhibition of histone phosphorylation by staurosporine leads to chromosome decondensation. J. Biol. Chem. 269, 9568–9573 (1994).
Chadee, D. N., Allis, C. D., Wright, J. A. & Davie, J. R. Histone H1b phosphorylation is dependent upon ongoing transcription and replication in normal and ras-transformed mouse fibroblasts. J. Biol. Chem. 272, 8113–8116 ( 1997).
Talasz, H., Sapojnikova, N., Helliger, W., Lindner, H. & Puschendorf, B. In vitro binding of H1 histone subtypes to nucleosomal organized mouse mammary tumor virus long terminal repeat promotor. J. Biol. Chem. 273, 32236 –32243 (1998).
Pruss, D. et al. An asymmetric model for the nucleosome: a binding site for linker histones inside the DNA gyres. Science 274, 614–617 (1996).
Roberge, M., Th’ng, J., Hamaguchi, J. & Bradbury, E. M. The topoisomerase II inhibitor VM-26 induces marked changes in histone H1 kinase activity, histones H1 and H3 phosphorylation, and chromosome condensation in G2 phase and mitotic BHK cells. J. Cell Biol. 111 , 1753–1762 (1990).
Paulson, J. R., Ciesielski, W. A., Schram, B. R. & Mesner, P. W. Okadaic acid induces dephosphorylation of histone H1 in metaphase-arrested HeLa cells. J. Cell Sci. 107, 267– 273 (1994).
Guo, X. W. et al. Chromosome condensation induced by fostriecin does not require p34cdc2 kinase activity and histone H1 hyperphosphorylation, but is associated with enhanced histone H2A and H3 phosphorylation. EMBO J. 14, 976–985 (1995).
Ajiro, K., Yoda, K., Utsumi, K. & Nishikawa, Y. Alteration of cell cycle-dependent histone phosphorylations by okadaic acid. Induction of mitosis-specific H3 phosphorylation and chromatin condensation in mammalian interphase cells. J. Biol. Chem. 271, 13197 –13201 (1996).
Bradbury, E. M. Reversible histone modifications and the chromosome cell cycle. BioEssays 14, 9–16 ( 1992).
Roth, S. Y. & Allis, C. D. Chromatin condensation: does histone H1 dephosphorylation play a role? Trends Biochem. Sci. 17, 93–98 (1992).
Lee, H. L. & Archer, T. K. Prolonged glucocorticoid exposure dephosphorylates histone H1 and inactivates the MMTV promoter. EMBO J. 17, 1454–1466 ( 1998).
Chadee, D. N. et al. Increased phosphorylation of histone H1 in mouse fibroblasts transformed with oncogenes or constitutively active mitogen-activated protein kinase kinase. J. Biol. Chem. 270, 20098 –20105 (1995).
Shen, X., Yu, L., Weir, J. W. & Gorovsky, M. A. Linker histones are not essential and affect chromatin condensation in vivo. Cell 82, 47–56 ( 1995).
Dou, Y., Mizzen, C. A., Abrams, M., Allis, C. D. & Gorovsky, M. A. Phosphorylation of linker histone H1 regulates gene expression in vivo by mimicking H1 removal. Mol. Cell 4, 641–647 ( 1999).
Eick, S., Nicolai, M., Mumberg, D. & Doenecke, D. Human H1 histones: conserved and varied sequence elements in two H1 subtype genes. Eur. J. Cell Biol. 49, 110–115 (1989).
Parseghian, M. H., Harris, D. A., Rishwain, D. R. & Hamkalo, B. A. Characterization of a set of antibodies specific for three human histone H1 subtypes. Chromosoma 103, 198– 208 (1994).
Acknowledgements
The authors would like to thank M. Fillion and D. McDonald for technical assistance; C. Lee for help with quantitative analysis; and T. Misteli for helpful discussions and critical reading of the manuscript. We thank L. Tourcotte and J. Turner for providing reagents and advice in energy-depletion experiments and M. Parseghian and B. Hamkalo for providing anti-histone H1 antibodies. This work was funded by the Alberta Cancer Foundation (M.J.H.) and the Medical Research Council of Canada (J.P.T. and M.J.H.). M.A.L. is supported by a scholarship from the Natural Sciences and Engineering Research Council of Canada, and M.J.H. is supported by a scholarship from the Medical Research Council of Canada.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Lever, M., Th'ng, J., Sun, X. et al. Rapid exchange of histone H1.1 on chromatin in living human cells. Nature 408, 873–876 (2000). https://doi.org/10.1038/35048603
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/35048603
This article is cited by
-
Release of linker histone from the nucleosome driven by polyelectrolyte competition with a disordered protein
Nature Chemistry (2022)
-
The solid and liquid states of chromatin
Epigenetics & Chromatin (2021)
-
Chromatin fibers stabilize nucleosomes under torsional stress
Nature Communications (2020)
-
Rett syndrome-causing mutations compromise MeCP2-mediated liquid–liquid phase separation of chromatin
Cell Research (2020)
-
Chromatin accessibility and the regulatory epigenome
Nature Reviews Genetics (2019)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.