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5' exon interactions within the human spliceosome establish a framework for exon junction complex structure and
Vienna L Reichert1, Hervé Le Hir, Melissa S Jurica
1Howard Hughes Medical Institute, Department of Biochemistry, Brandeis University, Massachusetts 02454, USA.
Genes & Development
|November 5, 2002
Summary
The exon junction complex (EJC) is assembled during pre-mRNA splicing. REF/Aly, Y14, and Magoh form the stable EJC core on spliced mRNA.
Area of Science:
- Molecular Biology
- RNA Processing
- Gene Expression Regulation
Background:
- Splicing deposits the exon junction complex (EJC) near exon-exon junctions on spliced mRNAs.
- The EJC plays roles in mRNA export, localization, and nonsense-mediated decay.
Purpose of the Study:
- To investigate the mechanism and timing of EJC assembly during pre-mRNA splicing.
- To identify the core components of the EJC that remain stably bound to mRNA.
Main Methods:
- Analysis of protein-RNA interactions during splicing.
- Mass spectrometry and Western blotting of purified complexes (H, C, and mRNP).
Main Results:
- Proteins interact dynamically with the 5' exon during splicing.
- REF/Aly binds pre-mRNA before spliceosome assembly.
- Y14, Magoh, RNPS1, UAP56, and SRm160 are found in spliceosomes.
- Upon ligation, RNPS1, UAP56, and SRm160 dissociate, while REF/Aly, Y14, and Magoh remain bound.
Conclusions:
- REF/Aly, Y14, and Magoh constitute the stable core of the exon junction complex.
- EJC assembly is a stepwise process occurring during and after splicing.