Coupled in vitro synthesis and splicing of RNA polymerase II transcripts.

Journal Article

Compelling in vivo studies suggest a tight functional linkage between RNA polymerase II transcription and premessenger RNA splicing. At present, the specific interactions involved in this coupling are poorly understood and deserve investigation. To this end, we developed an in vitro system that permits study of coupled transcription and splicing. Transcripts generated by RNA polymerase II were accurately and efficiently spliced under reaction conditions that permitted both transcription and splicing to occur simultaneously. The splicing of RNA-polymerase-II-driven transcripts was accelerated relative to that of the same transcripts driven by T7 RNA polymerase. Moreover, the product of exon ligation was found associated with the DNA template in reactions driven by RNA polymerase II. These two findings indicate that transcription and splicing were coupled in the in vitro system driven by RNA polymerase II, and suggest that this system will be useful for the biochemical study of this coupling.

Full Text

Duke Authors

Cited Authors

  • Ghosh, S; Garcia-Blanco, MA

Published Date

  • September 2000

Published In

Volume / Issue

  • 6 / 9

Start / End Page

  • 1325 - 1334

PubMed ID

  • 10999609

International Standard Serial Number (ISSN)

  • 1355-8382

Language

  • eng

Conference Location

  • United States