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PMID: 16651655 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Functional coupling of RNAP II transcription to spliceosome assembly.

Genes & development ·Vol. 20 ·No. 9 ·2006-05-01 ·Pages 1100-9

Das R, Dufu K, Romney B, Feldt M, Elenko M, Reed R

Abstract

The pathway of gene expression in higher eukaryotes involves a highly complex network of physical and functional interactions among the different machines involved in each step of the pathway. Here we established an efficient in vitro system to determine how RNA polymerase II (RNAP II) transcription is functionally coupled to pre-mRNA splicing. Strikingly, our data show that nascent pre-messenger RNA (pre-mRNA) synthesized by RNAP II is immediately and quantitatively directed into the spliceosome assembly pathway. In contrast, nascent pre-mRNA synthesized by T7 RNA polymerase is quantitatively assembled into the nonspecific H complex, which consists of heterogeneous nuclear ribonucleoprotein (hnRNP) proteins and is inhibitory for spliceosome assembly. Consequently, RNAP II transcription results in a dramatic increase in both the kinetics of splicing and overall yield of spliced mRNA relative to that observed for T7 transcription. We conclude that RNAP II mediates the functional coupling of transcription to splicing by directing the nascent pre-mRNA into spliceosome assembly, thereby bypassing interaction of the pre-mRNA with the inhibitory hnRNP proteins.

MeSH Terms
DNA-Directed RNA Polymerases/genetics,metabolism HeLa Cells Humans RNA Polymerase II/genetics,metabolism RNA Precursors/genetics,metabolism RNA Splicing Ribonucleoproteins/genetics,metabolism Spliceosomes/physiology Transcription, Genetic Viral Proteins/genetics,metabolism
Chemicals
RNA Precursors Ribonucleoproteins Viral Proteins RNA Polymerase II bacteriophage T7 RNA polymerase DNA-Directed RNA Polymerases
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Das Rita
Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, USA.
Dufu Kobina
Romney Ben
Feldt Megan
Elenko Mark
Reed Robin
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Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
0890-9369
DOI
10.1101/gad.1397406
Published
2006-05-01
Pages
1100-9
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC1472470
Subset
IM
Grants
NIGMS NIH HHS · R01 GM043375 · United States
External Links
PubMed source DOI: 10.1101/gad.1397406 PMC Full Text View journal details
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