Splicing remodels messenger ribonucleoprotein architecture via eIF4A3-dependent and -independent recruitment of exon junction complex components

Zhang, Z., Krainer, A. R. (July 2007) Splicing remodels messenger ribonucleoprotein architecture via eIF4A3-dependent and -independent recruitment of exon junction complex components. Proc Natl Acad Sci U S A, 104 (28). pp. 11574-9. ISSN 0027-8424 (Print)

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URL: http://www.ncbi.nlm.nih.gov/pubmed/17606899
DOI: 10.1073/pnas.0704946104

Abstract

Pre-mRNA splicing not only removes introns and joins exons to generate spliced mRNA but also results in remodeling of the spliced messenger ribonucleoprotein, influencing various downstream events. This remodeling includes the loading of an exon-exon junction complex (EJC). It is unclear how the spliceosome recruits the EJC onto the mRNA and whether EJC formation or EJC components are required for pre-mRNA splicing. Here we immunodepleted the EJC core component eIF4A3 from HeLa cell nuclear extract and found that eIF4A3 is dispensable for pre-mRNA splicing in vitro. However, eIF4A3 is required for the splicing-dependent loading of the Y14/Magoh heterodimer onto mRNA, and this activity of human eIF4A3 is also present in the Drosophila ortholog. Surprisingly, the loading of six other EJC components was not affected by eIF4A3 depletion, suggesting that their binding to mRNA involves different or redundant pathways. Finally, we found that the assembly of the EJC onto mRNA occurs at the late stages of the splicing reaction and requires the second-step splicing and mRNA-release factor HRH1/hPrp22. The EJC-dependent and -independent recruitment of RNA-binding proteins onto mRNA suggests a role for the EJC in messenger ribonucleoprotein remodeling involving interactions with other proteins already bound to the pre-mRNA, which has implications for nonsense-mediated mRNA decay and other mRNA transactions.

Item Type: Paper
Uncontrolled Keywords: Animals DEAD-box RNA Helicases/genetics/ physiology Drosophila Proteins/genetics/metabolism Exons/ genetics Humans Mice Nuclear Proteins/genetics/metabolism RNA Precursors/genetics/metabolism RNA Splice Sites/genetics RNA Splicing/ genetics RNA-Binding Proteins/genetics/metabolism Ribonucleoproteins/genetics/ metabolism
Subjects: organism description > animal > insect > Drosophila
organism description > animal
organism description > animal > mammal > primates > hominids > human
organism description > animal > mammal > rodent > mouse
CSHL Authors:
Communities: CSHL labs > Krainer lab
Depositing User: CSHL Librarian
Date: 10 July 2007
Date Deposited: 29 Aug 2011 23:17
Last Modified: 02 Apr 2014 16:19
PMCID: PMC1913901
Related URLs:
URI: http://repository.cshl.edu/id/eprint/15296

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