Dynamic binding of histone H1 to chromatin in living cells

The linker histone H1 is believed to be involved in chromatin organization by stabilizing higher-order chromatin structure. Histone H1 is generally viewed as a repressor of transcription as it prevents the access of transcription factors and chromatin remodelling complexes to DNA. Determining the binding properties of histone H1 to chromatin in vivo is central to understanding how it exerts these functions. We have used photobleaching techniques to measure the dynamic binding of histone H1–GFP to unperturbed chromatin in living cells. Here we show that almost the entire population of H1–GFP is bound to chromatin at any one time; however, H1–GFP is exchanged continuously between chromatin regions. The residence time of H1–GFP on chromatin between exchange events is several minutes in both euchromatin and heterochromatin. In addition to the mobile fraction, we detected a kinetically distinct, less mobile fraction. After hyperacetylation of core histones, the residence time of H1–GFP is reduced, suggesting a higher rate of exchange upon chromatin remodelling. These results support a model in which linker histones bind dynamically to chromatin in a stop-and-go mode.

[1]  J. Davie,et al.  Histone acetylation alters the capacity of the H1 histones to condense transcriptionally active/competent chromatin. , 1990, The Journal of biological chemistry.

[2]  P. Becker,et al.  Reconstitution of hyperacetylated, DNase I-sensitive chromatin characterized by high conformational flexibility of nucleosomal DNA. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[3]  L. M. Lira,et al.  Sequence-specific antirepression of histone H1-mediated inhibition of basal RNA polymerase II transcription. , 1991, Science.

[4]  A. Houtsmuller,et al.  Action of DNA repair endonuclease ERCC1/XPF in living cells. , 1999, Science.

[5]  K. V. van Holde,et al.  Linker histones versus HMG1/2: a struggle for dominance? , 1998, BioEssays : news and reviews in molecular, cellular and developmental biology.

[6]  J. O. Thomas,et al.  Histone H1: location and role. , 1999, Current opinion in cell biology.

[7]  M. Grunstein.  Histone acetylation in chromatin structure and transcription , 1997, Nature.

[8]  M. Vettese-Dadey,et al.  Differential repression of transcription factor binding by histone H1 is regulated by the core histone amino termini. , 1994, The EMBO journal.

[9]  V. Ramakrishnan,et al.  Histone H1 and chromatin higher-order structure. , 1997, Critical reviews in eukaryotic gene expression.

[10]  K. Struhl.  Histone acetylation and transcriptional regulatory mechanisms. , 1998, Genes & development.

[11]  S. Berger.  Gene activation by histone and factor acetyltransferases. , 1999, Current opinion in cell biology.

[12]  David T. Brown,et al.  Differential effect of H1 variant overexpression on cell cycle progression and gene expression. , 1996, Nucleic acids research.

[13]  T. Misteli,et al.  High mobility of proteins in the mammalian cell nucleus , 2000, Nature.

[14]  M. Rechsteiner,et al.  Dynamic behavior of histone H1 microinjected into HeLa cells , 1986, The Journal of cell biology.

[15]  F. Thoma,et al.  Influence of histone H1 on chromatin structure , 1977, Cell.

[16]  T. R. Hebbes,et al.  A direct link between core histone acetylation and transcriptionally active chromatin. , 1988, The EMBO journal.

[17]  C. Allis,et al.  Phosphorylated and dephosphorylated linker histone H1 reside in distinct chromatin domains in Tetrahymena macronuclei. , 1995, Molecular biology of the cell.

[18]  T. Misteli,et al.  Serine/threonine phosphatase 1 modulates the subnuclear distribution of pre-mRNA splicing factors. , 1996, Molecular biology of the cell.

[19]  H. Richard-Foy,et al.  The transcriptionally-active MMTV promoter is depleted of histone H1. , 1992, Nucleic acids research.

[20]  R. Chalkley,et al.  Exchange of histones H1, H2A, and H2B in vivo. , 1985, Biochemistry.

[21]  David T. Brown,et al.  Effects of H1 Histone Variant Overexpression on Chromatin Structure* , 1999, The Journal of Biological Chemistry.

[22]  A. Wolffe,et al.  Core histone acetylation does not block linker histone binding to a nucleosome including a Xenopus borealis 5 S rRNA gene. , 1994, The Journal of biological chemistry.

[23]  A. Gunjan,et al.  Overproduction of histone H1 variants in vivo increases basal and induced activity of the mouse mammary tumor virus promoter. , 1999, Nucleic acids research.

[24]  J. O. Thomas,et al.  Exchange of histone H1 between segments of chromatin. , 1981, Journal of molecular biology.

[25]  D. Bates,et al.  Stability of the higher-order structure of chicken-erythrocyte chromatin in solution. , 1981, European journal of biochemistry.

[26]  C. Allis,et al.  Phosphorylation of linker histone H1 regulates gene expression in vivo by mimicking H1 removal. , 1999, Molecular cell.

[27]  A. Murray,et al.  Interphase chromosomes undergo constrained diffusional motion in living cells , 1997, Current Biology.

[28]  C. Allis,et al.  The language of covalent histone modifications , 2000, Nature.

[29]  Y. Allory,et al.  Localization and phosphorylation of HP1 proteins during the cell cycle in mammalian cells , 1999, Chromosoma.