Mapping interactions between mRNA export factors in living cells

The TREX complex couples nuclear mRNA processing events with subsequent export to the cytoplasm. TREX also acts as a binding platform for the mRNA export receptor Nxf1. The sites of mRNA transcription and processing within the nucleus have been studied extensively. However, little is known about whe...

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Bibliographic Details
Published in:PloS one 2013-06, Vol.8 (6), p.e67676-e67676
Main Authors: Teng, I-Fang, Wilson, Stuart A
Format: Article
Language:English
Subjects:
Assembly
Biology
Cell Nucleus - metabolism
Cell Survival
Cells (biology)
Complex formation
Cytoplasm
Energy transfer
Exports
Fluorescence
Fluorescence Recovery After Photobleaching
Fluorescence Resonance Energy Transfer
Gene expression
HeLa Cells
Humans
Immunoglobulins
Molecular biology
mRNA processing
Nuclear Proteins - metabolism
Nuclei
Nucleocytoplasmic Transport Proteins - metabolism
Protein Binding
Protein Interaction Mapping
Protein Transport
Proteins
Recovery (Medical)
RNA polymerase
RNA Transport
RNA-Binding Proteins - metabolism
Splicing
Transcription
Transcription Factors - metabolism
Transcription, Genetic
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Summary:The TREX complex couples nuclear mRNA processing events with subsequent export to the cytoplasm. TREX also acts as a binding platform for the mRNA export receptor Nxf1. The sites of mRNA transcription and processing within the nucleus have been studied extensively. However, little is known about where TREX assembly takes place and where Nxf1 is recruited to TREX to form the export competent mRNP. Here we have used sensitized emission Förster resonance energy transfer (FRET) and fluorescence lifetime imaging (FLIM)-FRET, to produce a spatial map in living cells of the sites for the interaction of two TREX subunits, Alyref and Chtop, with Nxf1. Prominent assembly sites for export factors are found in the vicinity of nuclear speckles in regions known to be involved in transcription, splicing and exon junction complex formation highlighting the close coupling of mRNA export with mRNP biogenesis.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0067676