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First published online 29 November 2005
doi: 10.1242/jcs.02689
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Research Article |
1 Division of Molecular Genetics, Deutsches Krebsforschungszentrum, Im Neuenheimer Feld 280, 69120 Heidelberg, Germany
2 Molecular Biophysics Group, Kirchhoff-Institut für Physik, Im Neuenheimer Feld 227, 69120 Heidelberg, Germany
Author for correspondence (e-mail: karsten.rippe{at}kip.uni-heidelberg.de)
Accepted 12 September 2005
In eukaryotes, the interaction of DNA with proteins and supramolecular complexes involved in gene expression is controlled by the dynamic organization of chromatin inasmuch as it defines the DNA accessibility. Here, the nuclear distribution of microinjected fluorescein-labeled dextrans of 42 kDa to 2.5 MDa molecular mass was used to characterize the chromatin accessibility in dependence on histone acetylation. Measurements of the fluorescein-dextran sizes were combined with an image correlation spectroscopy analysis, and three different interphase chromatin condensation states with apparent pore sizes of 16-20 nm, 36-56 nm and 60-100 nm were identified. A reversible change of the chromatin conformation to a uniform 60-100 nm pore size distribution was observed upon increased histone acetylation. This result identifies histone acetylation as a central factor in the dynamic regulation of chromatin accessibility during interphase. In mitotic chromosomes, the chromatin exclusion limit was 10-20 nm and independent of the histone acetylation state.
Key words: Heterochromatin, Image correlation spectroscopy, Trichostatin A, Microinjection
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