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Mol Cell. 2010 Nov 24;40(4):582-93. doi: 10.1016/j.molcel.2010.11.005.

Splicing-dependent RNA polymerase pausing in yeast.

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  • 1Wellcome Trust Centre for Cell Biology, University of Edinburgh, King's Buildings, Edinburgh EH9 3JR, UK.

Abstract

In eukaryotic cells, there is evidence for functional coupling between transcription and processing of pre-mRNAs. To better understand this coupling, we performed a high-resolution kinetic analysis of transcription and splicing in budding yeast. This revealed that shortly after induction of transcription, RNA polymerase accumulates transiently around the 3' end of the intron on two reporter genes. This apparent transcriptional pause coincides with splicing factor recruitment and with the first detection of spliced mRNA and is repeated periodically thereafter. Pausing requires productive splicing, as it is lost upon mutation of the intron and restored by suppressing the splicing defect. The carboxy-terminal domain of the paused polymerase large subunit is hyperphosphorylated on serine 5, and phosphorylation of serine 2 is first detected here. Phosphorylated polymerase also accumulates around the 3' splice sites of constitutively expressed, endogenous yeast genes. We propose that transcriptional pausing is imposed by a checkpoint associated with cotranscriptional splicing.

Copyright © 2010 Elsevier Inc. All rights reserved.

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  • A pause to splice. [Mol Cell. 2010]
PMID:
21095588
[PubMed - indexed for MEDLINE]
PMCID:
PMC3000496
Free PMC Article
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