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An RNA polymerase has been thought to transcribe by seeking out a promoter, initiating and then tracking down the template. We add tumor necrosis factor α to primary human cells, switch on transcription of a 221-kb gene and monitor promoter position during the ensuing transcription cycle (using RNA fluorescence in situ hybridization coupled to super-resolution localization, chromosome conformation capture and Monte Carlo simulations). Results are consistent with a polymerase immobilized in a 'factory' capturing a promoter and reeling in the template, as the transcript and promoter are extruded. Initially, the extruded promoter is tethered close to the factory and so likely to re-initiate; later, the tether becomes long enough to allow re-initiation in another factory. We suggest close tethering underlies enhancer function and transcriptional 'bursting'.

Original publication

DOI

10.1093/nar/gks1441

Type

Journal article

Journal

Nucleic Acids Res

Publication Date

01/02/2013

Volume

41

Pages

2216 - 2227

Keywords

Cells, Cultured, DNA-Directed RNA Polymerases, Human Umbilical Vein Endothelial Cells, Humans, In Situ Hybridization, Fluorescence, Models, Genetic, Monte Carlo Method, Promoter Regions, Genetic, RNA, Repressor Proteins, Templates, Genetic, Transcription, Genetic