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- Title
Cotranscriptional coupling of splicing factor recruitment and precursor messenger RNA splicing in mammalian cells.
- Authors
Listerman, Imke; Sapra, Aparna K.; Neugebauer, Karla M.
- Abstract
Coupling between transcription and RNA processing is a key gene regulatory mechanism. Here we use chromatin immunoprecipitation to detect transcription-dependent accumulation of the precursor mRNA (pre-mRNA) splicing factors hnRNP A1, U2AF65 and U1 and U5 snRNPs on the intron-containing human FOS gene. These factors were poorly detected on intronless heat-shock and histone genes, a result that opposes direct recruitment by RNA polymerase II (Pol II) or the cap-binding complex in vivo. However, an observed RNA-dependent interaction between U2AF65 and active forms of Pol II may stabilize U2AF65 binding to intron-containing nascent RNA. We establish chromatin-RNA immunoprecipitation and show that FOS pre-mRNA is cotranscriptionally spliced. Notably, the topoisomerase I inhibitor camptothecin, which stalls elongating Pol II, increased cotranscriptional splicing factor accumulation and splicing in parallel. This provides direct evidence for a kinetic link between transcription, splicing factor recruitment and splicing catalysis.
- Subjects
RNA splicing; GENE expression; CARRIER proteins; DROSOPHILA melanogaster; CHROMATIN
- Publication
Nature Structural & Molecular Biology, 2006, Vol 13, Issue 9, p815
- ISSN
1545-9993
- Publication type
Article
- DOI
10.1038/nsmb1135