Binding of the RNA polymerase I transcription complex to its promoter can modify positioning of downstream nucleosomes assembled in vitro

Philippe Georgel, Borries Demeler, Chris Terpening, Marvin R. Paule, Kensal E. Van Holde

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

We have studied the reconstitution of chromatin-like structures in vitro, using purified RNA polymerase I transcription complexes and histone octamers. The plasmid construct used in these studies is a pUC8 derivative in which we have inserted an RNA polymerase I core promoter region of Acanthamoeba castellanii upstream of four repeats of the 5 S rDNA nucleosome positioning sequence (208 base pairs) from Lytechinus variegatus. When histone octamers were reconstituted onto the naked DNA template, the expected nucleosome positioning previously observed using tandem repeats of the same 208-base pair fragment was not obtained (as assayed by restriction enzyme digestion mapping of the inserted region of the plasmid). We show that the location of the RNA polymerase I core promoter region with regard to the tandemly repeated 208-base pair positioning sequence is a major determinant in the positioning of the histone octamers. Reconstituting first with the stalled transcription complex excluded octamers from the promoter region and recovered the expected nucleosome positioning downstream on the four repeats of the 5 S positioning sequence. The observed competition between histone octamers and the transcription complex for the promoter region suggests a great similarity with what has been reported from in vitro studies of RNA polymerase II and III transcription systems. We may be looking at a mechanism of regulation of transcription for the RNA polymerase I.

Original languageEnglish (US)
Pages (from-to)1947-1954
Number of pages8
JournalJournal of Biological Chemistry
Volume268
Issue number3
StatePublished - Jan 25 1993
Externally publishedYes

Fingerprint

RNA Polymerase I
Nucleosomes
Transcription
Genetic Promoter Regions
Histones
Base Pairing
Plasmids
Lytechinus
Acanthamoeba castellanii
RNA Polymerase III
Restriction Mapping
Tandem Repeat Sequences
RNA Polymerase II
Ribosomal DNA
Chromatin
Digestion
In Vitro Techniques
DNA
Derivatives
Enzymes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Binding of the RNA polymerase I transcription complex to its promoter can modify positioning of downstream nucleosomes assembled in vitro. / Georgel, Philippe; Demeler, Borries; Terpening, Chris; Paule, Marvin R.; Van Holde, Kensal E.

In: Journal of Biological Chemistry, Vol. 268, No. 3, 25.01.1993, p. 1947-1954.

Research output: Contribution to journalArticle

Georgel, Philippe ; Demeler, Borries ; Terpening, Chris ; Paule, Marvin R. ; Van Holde, Kensal E. / Binding of the RNA polymerase I transcription complex to its promoter can modify positioning of downstream nucleosomes assembled in vitro. In: Journal of Biological Chemistry. 1993 ; Vol. 268, No. 3. pp. 1947-1954.
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