Journal
EMBO JOURNAL
Volume 23, Issue 8, Pages 1782-1791Publisher
WILEY
DOI: 10.1038/sj.emboj.7600187
Keywords
p54(nrb); PSF; RNA polymerase II; splicing complexes; transcription
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Funding
- NIGMS NIH HHS [GM 49044, R01 GM049044] Funding Source: Medline
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The functional coupling of transcription and splicing has been reported both in vivo and in vitro, but the molecular mechanisms governing these interactions remain largely unknown. Here we show that p54(nrb), a transcription/splicing factor, associates with the 5'splice site (SS) within large complexes present in HeLa cell nuclear extracts, in which the hyperphosphorylated form of RNA polymerase II (RNAPIIO) is associated with U1 or U1 and U2 snRNPs. These RNAPIIO-snRNP complexes also contain other transcription/splicing factors, such as PSF and TLS, as well as transcription factors that interact with RNAPIIO during elongation, including P-TEFb, TAT-SF1 and TFIIF. The presence of these factors in functional elongation complexes, demonstrated using an immobilized DNA template assay, strongly suggests that the RNAPIIO-snRNP complexes reflect physiologically relevant interactions between the transcription and splicing machineries. Our finding that both p54(nrb) and PSF, which bind the C-terminal domain of the largest subunit of RNAPII, can interact directly with the 5'SS indicates that these factors may mediate contacts between RNAPII and snRNPs during the coupled transcription/splicing process.
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