Fate of linear and supercoiled multinucleosomic templates during transcription.

Details

Serval ID
serval:BIB_742E8D2F31B7
Type
Article: article from journal or magazin.
Collection
Publications
Title
Fate of linear and supercoiled multinucleosomic templates during transcription.
Journal
EMBO Journal
Author(s)
ten Heggeler-Bordier B., Schild-Poulter C., Chapel S., Wahli W.
ISSN
0261-4189[print], 0261-4189[linking]
Publication state
Published
Issued date
06/1995
Volume
14
Number
11
Pages
2561-2569
Language
english
Notes
Publication types: In Vitro ; Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
Abstract
Electron microscopy was used to monitor the fate of reconstituted nucleosome cores during in vitro transcription of long linear and supercoiled multinucleosomic templates by the prokaryotic T7 RNA polymerase and the eukaryotic RNA polymerase II. Transcription by T7 RNA polymerase disrupted the nucleosomal configuration in the transcribed region, while nucleosomes were preserved upstream of the transcription initiation site and in front of the polymerase. Nucleosome disruption was independent of the topology of the template, linear or supercoiled, and of the presence or absence of nucleosome positioning sequences in the transcribed region. In contrast, the nucleosomal configuration was preserved during transcription from the vitellogenin B1 promoter with RNA polymerase II in a rat liver total nuclear extract. However, the persistence of nucleosomes on the template was not RNA polymerase II-specific, but was dependent on another activity present in the nuclear extract. This was demonstrated by addition of the extract to the T7 RNA polymerase transcription reaction, which resulted in retention of the nucleosomal configuration. This nuclear activity, also found in HeLa cell nuclei, is heat sensitive and could not be substituted by nucleoplasmin, chromatin assembly factor (CAF-I) or a combination thereof. Altogether, these results identify a novel nuclear activity, called herein transcription-dependent chromatin stabilizing activity I or TCSA-I, which may be involved in a nucleosome transfer mechanism during transcription.
Keywords
Animals, Cell Nucleus/metabolism, DNA, Superhelical/genetics, DNA, Superhelical/metabolism, DNA-Directed RNA Polymerases/metabolism, Hela Cells, Humans, Liver/metabolism, Microscopy, Electron, Nuclear Proteins/metabolism, Nucleoplasmins, Nucleosomes/metabolism, Nucleosomes/ultrastructure, Phosphoproteins, Plasmids/genetics, Plasmids/metabolism, RNA Polymerase II/metabolism, Rats, Transcription, Genetic, Viral Proteins
Pubmed
Web of science
Create date
24/01/2008 16:05
Last modification date
20/08/2019 14:31
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