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Title: Mapping mRNA export complex formation in living cells
Author: Teng, If
ISNI:       0000 0004 2745 1709
Awarding Body: University of Sheffield
Current Institution: University of Sheffield
Date of Award: 2013
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The TREX ('transcription/export') complex couples nuclear RNA processing events and undergoes substantial rearrangements with mRNA export receptor Nxf1 during assembly and maturation for efficient mRNA export. However, it is not clear 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 interactions of two TREX subunits, Alyref and Chtop, with Nxf1. In addition, we have performed fluorescence recovery after photobleaching (FRAP) to monitor the dynamic behaviors of export factors in the nucleus, showing Chotp and Alyref have distinctive attributes between speckle and nucleoplasm. Together, our results have been shown that 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.
Supervisor: Wilson, Stuart Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available