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J Biol Chem, Vol. 273, Issue 29, 18028-18039, July 17, 1998
From the Dipartimento di Biologia Stutterale e Funzionale III
Facoltá di Scienze, Universitá di Milano,
21100 Varese, Italy
We report the characterization of an in
vitro chromatin assembly system derived from Artemia
embryos and its application to the study of AluI-113
satellite DNA organization in nucleosomes. The system efficiently
reconstitutes chromatin templates by associating DNA, core histones,
and H1. The polynucleosomal complexes show physiological spacing of
repeat length 190 ± 5 base pairs, and the internucleosomal
distances are modulated by energy-using activities that contribute to
the dynamics of chromatin conformation. The assembly extract was used
to reconstitute tandemly repeated AluI-113 sequences. The
establishment of preferred histone octamer/satellite DNA interactions
was observed. In vitro, AluI-113 elements
dictated the same nucleosome translational localizations as found
in vivo. Specific rotational constraints seem to be the
central structural requirement for nucleosome association. Satellite
dinucleosomes showed decreased translational mobility compared with
mononucleosomes. This could be the consequence of interactions between
rotationally positioned nucleosomes separated by linker DNA of uniform
length. AluI-113 DNA led to weak cooperativity of
nucleosome association in the proximal flanking regions, which
decreased with distance. Moreover, the structural properties of
satellite chromatin can spread, thus leading to a specific organization
of adjacent nucleosomes.
In Vitro Reconstitution of Artemia
Satellite Chromatin
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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