Dynamic binding of histone H1 to chromatin in living cells

Nature. 2000 Dec 14;408(6814):877-81. doi: 10.1038/35048610.

Abstract

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.

MeSH terms

  • 3T3 Cells
  • Acetylation
  • Animals
  • Cell Line
  • Chromatin / metabolism*
  • Chromatography, High Pressure Liquid
  • Green Fluorescent Proteins
  • Heterochromatin / metabolism
  • Histones / metabolism*
  • Luminescent Proteins / metabolism
  • Mice
  • Mice, Inbred C57BL
  • Microscopy, Fluorescence
  • Protein Binding
  • Recombinant Fusion Proteins / metabolism

Substances

  • Chromatin
  • Heterochromatin
  • Histones
  • Luminescent Proteins
  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins