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Mol. Cell. Biol., 10 1995, 5294-5303, Vol 15, No. 10
R Dammann, R Lucchini, T Koller and JM Sogo
In growing yeast cells, about half of the 150 tandemly repeated rRNA genes
are transcriptionally active and devoid of nucleosomes. By using the
intercalating drug psoralen as a tool to mark accessible sites along
chromatin DNA in vivo, we found that the active rRNA gene copies are rather
randomly distributed along the ribosomal rRNA gene locus. Moreover, results
from the analysis of a single, tagged transcription unit in the tandem
array are not consistent with the presence of a specific subset of active
genes that is stably maintained throughout cell divisions. In the rRNA
intergenic spacers of yeast cells, an enhancer is located at the 3' end of
each transcription unit, 2 kb upstream of the next promoter. Analysis of
the chromatin structure along the tandem array revealed a structural link
between transcription units and adjacent, 3' flanking enhancer sequences:
each transcriptionally active gene is flanked by a nonnucleosomal enhancer,
whereas inactive, nucleosome-packed gene copies are followed by enhancers
regularly packaged in nucleosomes. From the fact that nucleosome-free
enhancers were also detected in an RNA polymerase I mutant strain, we
interpret these open chromatin structures as being the result of specific
protein-DNA interactions that can occur before the onset of transcription.
In contrast, in this mutant strain, all of the rRNA coding sequences are
packaged in nucleosomal arrays. This finding indicates that the
establishment of the open chromatin conformation on the activated gene
copies requires elongating RNA polymerase I molecules advancing through the
template.
Copyright © 1995, American Society for Microbiology
Transcription in the yeast rRNA gene locus: distribution of the active gene copies and chromatin structure of their flanking regulatory sequences
Institute of Cell Biology, Eidgenossiche Technische Hochschule- Honggerberg, Zurich, Switzerland.
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