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Nucleic Acids Research, 1994, Vol. 22, No. 6 937-945
© 1994

MOLECULAR BIOLOGY

Octamer displacement and redistribution in transcription of single nucleosomes

Marie-Françoise O'Donohue, Isabelle Duband-Goulet, Ali Hamiche and Ariel Prunell*

Institut Jacques Monod, Centre National de la Recherche Scientifique et Université Paris 7 2 place Jussieu, 75251 Paris Cédex 05, France

*To whom correspondence should be addressed

Received January 10, 1994. Accepted February 21, 1994.

Single nucleosomes were assembled on a 357bp DNA fragment containing a 5S RNA gene from sea urchin and a promoter for SP6 RNA polymerase, and were fractionated as a function of their positions by gel electrophoresls (1,2). Transcribed nucleosome positions were detected by observing band disappearance in gels, which In turn provided evidence for the displacement of the histone octamer upon transcription. Differential band disappearance showed that nucleosomes closer to the promoter were harder to transcribe, and transcription was blocked when the nucleosome proximal boundary was at the start site. Nucleosomes located at discrete positions were also eluted from the gel bands and transcribed. In this case, new bands appeared as a consequence of octamer redistribution. Such redistribution occurred over all untranscrlbed positions, as well as over transcribed positions close enough to the promoter. Similar conclusions were derived from another previously investigated fragment containing a Xenopus 5S RNA gene (3,4).


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