Molecular Cell
Volume 40, Issue 4, 24 November 2010, Pages 582-593
Journal home page for Molecular Cell

Article
Splicing-Dependent RNA Polymerase Pausing in Yeast

https://doi.org/10.1016/j.molcel.2010.11.005Get rights and content
Under a Creative Commons license
open access

Summary

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.

Highlights

► RNA polymerase II pauses at the 3′ end of an intron ► RNA polymerase II pausing depends on productive splicing ► Dynamic phosphorylation of polymerase changes according to intron splicing status ► We propose a transcriptional checkpoint that responds to cotranscriptional splicing

Cited by (0)