RNA catalyses nuclear pre-mRNA splicing

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RNAcatalyses nuclear pre-mRNA splicing

In nuclear pre-messenger RNA splicing, introns are excised by the spliceosome, a dynamic machine composed of both proteins and small nuclear RNAs (snRNAs). Over thirty years ago, after the discovery of self-splicing group II intron RNAs, the snRNAswere proposed to catalyse splicing. However, no definitive evidence for a role of either RNAor protein in catalysis by the spliceosome has been repor...

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TRAP150 activates pre-mRNA splicing and promotes nuclear mRNA degradation

TRAP150 has been identified as a subunit of the transcription regulatory complex TRAP/Mediator, and also a component of the spliceosome. The exact function of TRAP150, however, remains unclear. We recently identified TRAP150 by its association with the mRNA export factor TAP. TRAP150 contains an arginine/serine-rich domain and has sequence similarity with the cell death-promoting transcriptiona...

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Analysis of pre-mRNA splicing using HeLa cell nuclear extracts.

This protocol is used to determine the splicing behavior of pre-mRNAs in cell extracts that are capable of carrying out splicing (e.g., nuclear extracts from HeLa cells). (32)P-labeled RNA is incubated under splicing conditions for various times, and the resulting products are analyzed on denaturing polyacrylamide gels.

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Phosphorylated RNA polymerase II stimulates pre-mRNA splicing.

RNA polymerase II (RNAP II) is responsible for transcription of mRNA precursors in eukaryotic cells. Recent studies, however, have suggested that RNAP II also participates in subsequent RNA processing reactions through interactions between the carboxy-terminal domain (CTD) of the RNAP II largest subunit and processing factors. Using reconstituted in vitro splicing assays, we show that RNAP II f...

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Preparation of Splicing Competent Nuclear Extract from Mammalian Cells and In Vitro Pre-mRNA Splicing Assay.

The ability to perform in vitro splicing assays has paved the way for in-depth studies of the mechanisms and machinery involved in the process of splicing. The in vitro splicing assay is a valuable experimental approach that combines the complexity of the spliceosome and regulatory systems with the flexibility of performing endless splicing and alternative splicing reactions. Through the use of...

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ژورنال

عنوان ژورنال: Nature

سال: 2013

ISSN: 0028-0836,1476-4687

DOI: 10.1038/nature12734