Exon Junction Complexes Mediate the Enhancing Effect of Splicing on mRNA Expression

Intron-containing genes are generally expressed more effectively in human cells than are intronless versions of the same gene. We have asked whether this effect is due directly to splicing or instead reflects the action of components of the exon junction complex (EJC) that is assembled at splice jun...

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Published inProceedings of the National Academy of Sciences - PNAS Vol. 100; no. 20; pp. 11327 - 11332
Main Authors Wiegand, Heather L., Lu, Shihua, Cullen, Bryan R.
Format Journal Article
LanguageEnglish
Published United States National Academy of Sciences 30.09.2003
National Acad Sciences
Subjects
Base Sequence
Biochemistry
Biological Sciences
Cell Line
Cloning, Molecular
Comparative analysis
DNA Primers
Exons
Gene expression
Genes
Genomics
Humans
Introns
Messenger RNA
Plasmids
Protein synthesis
Ribonucleic acid
RNA
RNA Splicing
RNA, Messenger - genetics
Splicing
Untranslated regions
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Summary:Intron-containing genes are generally expressed more effectively in human cells than are intronless versions of the same gene. We have asked whether this effect is due directly to splicing or instead reflects the action of components of the exon junction complex (EJC) that is assembled at splice junctions after splicing is completed. Here, we show that intron removal does not enhance gene expression if EJC formation is blocked. Conversely, RNA tethering of the EJC components SRm160 or RNPS1 boosts the expression of intronless mRNAs but not of spliced mRNAs. Splicing and RNPS1 tethering are shown to enhance the same steps in mRNA biogenesis and function, including mRNA 3′ end processing and translation. Together, these data argue that the EJC is primarily responsible for the positive effect of splicing on gene expression.
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Communicated by Wolfgang K. Joklik, Duke University Medical Center, Durham, NC, August 1, 2003
H.L.W. and S.L. contributed equally to this work.
To whom correspondence should be addressed. E-mail: culle002@mc.duke.edu.
Abbreviations: CAT, chloramphenicol acetyltransferase; CMV, cytomegalovirus; EJC, exon junction complex; β-gal, β-galactosidase; RPA, RNase protection assay; SX, short exon.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.1934877100