Nucleic Acids Research, Vol 25, Issue 3 511-517, Copyright © 1997 by Oxford University Press
G Langst, T Schatz, J Langowski and I Grummt
We have analyzed the chromatin structure of mouse ribosomal RNA genes
(rDNA) by partial digestion of genomic DNA with micrococcal nuclease
(MNase), DNase I and identified hypersensitive sites by indirect end-
labeling. This analysis has revealed defined regions of nuclease
hypersensitivity in the intergenic spacer which in turn coincide with
regulatory elements. Hypersensitive sites map to the transcription
initiation site, the enhancer repeats, the spacer promoter and two sequence
elements which coincide with amplification-promoting sequences. Analysis of
the DNA curvature by computer modeling uncovered a striking correlation
between sequence-directed structural features of regulatory regions and the
position of nuclease hypersensitive sites. Moreover, we demonstrate that
nucleosomes are specifically positioned upstream and downstream of the
transcription start site. In vitro studies using chromatin assembled in the
presence of Drosophila embryo extracts show that binding of the
transcription termination factor TTF- I to the upstream terminator mediates
this specific nucleosome positioning at the rDNA promoter in an ATP-
dependent fashion.
ARTICLES
Structural analysis of mouse rDNA: coincidence between nuclease hypersensitive sites, DNA curvature and regulatory elements in the intergenic spacer
German Cancer Research Center, Division of Molecular Biology of the Cell II, 69120 Heidelberg, Germany.
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