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Genome-wide studies reveal that transcription by RNA polymerase II (Pol II) is dynamically regulated. To obtain a comprehensive view of a single transcription cycle, we switched on transcription of five long human genes (>100 kbp) with tumor necrosis factor-alpha (TNFalpha) and monitored (using microarrays, RNA fluorescence in situ hybridization, and chromatin immunoprecipitation) the appearance of nascent RNA, changes in binding of Pol II and two insulators (the cohesin subunit RAD21 and the CCCTC-binding factor CTCF), and modifications of histone H3. Activation triggers a wave of transcription that sweeps along the genes at approximately 3.1 kbp/min; splicing occurs cotranscriptionally, a major checkpoint acts several kilobases downstream of the transcription start site to regulate polymerase transit, and Pol II tends to stall at cohesin/CTCF binding sites.

Original publication

DOI

10.1073/pnas.0902573106

Type

Journal article

Journal

Proc Natl Acad Sci U S A

Publication Date

27/10/2009

Volume

106

Pages

18357 - 18361

Keywords

Binding Sites, CCCTC-Binding Factor, Cell Cycle Proteins, Cells, Cultured, Chromatin Immunoprecipitation, Chromosomal Proteins, Non-Histone, Humans, In Situ Hybridization, Introns, Nuclear Proteins, Oligonucleotide Array Sequence Analysis, Phosphoproteins, RNA Polymerase II, RNA Splicing, RNA, Messenger, Repressor Proteins, Transcription, Genetic, Transcriptional Activation, Tumor Necrosis Factor-alpha