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PMID: 12093754 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

The exon junction complex is detected on CBP80-bound but not eIF4E-bound mRNA in mammalian cells: dynamics of mRNP remodeling.

The EMBO journal ·Vol. 21 ·No. 13 ·2002-07-01 ·Pages 3536-45

Lejeune F, Ishigaki Y, Li X, Maquat LE

Abstract

Newly spliced mRNAs in mammalian cells are characterized by a complex of proteins at exon-exon junctions. This complex recruits Upf3 and Upf2, which function in nonsense-mediated mRNA decay (NMD). Both Upf proteins are detected on mRNA bound by the major nuclear cap-binding proteins CBP80/CBP20 but not mRNA bound by the major cytoplasmic cap-binding protein eIF4E. These and other data indicate that NMD targets CBP80-bound mRNA during a 'pioneer' round of translation, but whether nuclear eIF4E also binds nascent but dead-end transcripts is unclear. Here we provide evidence that nuclear CBP80 but not nuclear eIF4E is readily detected in association with intron-containing RNA and the C-terminal domain of RNA polymerase II. Consistent with this evidence, we demonstrate that RNPS1, Y14, SRm160, REF/Aly, TAP, Upf3X and Upf2 are detected in the nuclear fraction on CBP80-bound but not eIF4E-bound mRNA. Each of these proteins is also detected on CBP80-bound mRNA in the cytoplasmic fraction, indicating a presence on mRNA after export. The dynamics of mRNP composition before and after mRNA export are discussed.

MeSH Terms
Animals Antigens, Nuclear Antigens, Polyomavirus Transforming/genetics Biological Transport COS Cells/metabolism Chlorocebus aethiops Cytoplasm/metabolism DNA-Binding Proteins/metabolism Eukaryotic Initiation Factor-4E Exons/genetics Globins/genetics Introns/genetics Macromolecular Substances Nuclear Matrix-Associated Proteins Nuclear Proteins/metabolism Peptide Initiation Factors/metabolism Protein Interaction Mapping Protein Structure, Tertiary RNA Cap-Binding Proteins RNA Caps/metabolism RNA Polymerase II/metabolism RNA Precursors/metabolism RNA Processing, Post-Transcriptional RNA, Messenger/metabolism RNA-Binding Proteins/metabolism Ribonucleoproteins Simian virus 40/genetics Transcription Factors/metabolism Transfection
Chemicals
Antigens, Nuclear Antigens, Polyomavirus Transforming DNA-Binding Proteins Eukaryotic Initiation Factor-4E Macromolecular Substances Nuclear Matrix-Associated Proteins Nuclear Proteins Peptide Initiation Factors RBM8A protein, human RNA Cap-Binding Proteins RNA Caps RNA Precursors RNA, Messenger RNA-Binding Proteins RNPS1 protein, human Ribonucleoproteins SRRM1 protein, human Transcription Factors Globins RNA Polymerase II
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Lejeune Fabrice
Department of Biochemistry and Biophysics, School of Medicine and Dentistry, 601 Elmwood Avenue, Box 712, University of Rochester, Rochester, NY 14642, USA.
Ishigaki Yasuhito
Li Xiaojie
Maquat Lynne E
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/cdf345
Published
2002-07-01
Pages
3536-45
Language
English
Region
England
NLM ID
8208664
PMCID
PMC126094
Subset
IM
Grants
NIDDK NIH HHS · R01 DK033938 · United States
NIGMS NIH HHS · R01 GM059614 · United States
NIDDK NIH HHS · DK 33938 · United States
NIGMS NIH HHS · GM 59614 · United States
External Links
PubMed source DOI: 10.1093/emboj/cdf345 PMC Full Text View journal details
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