Abstract
Aberrant mRNAs with premature translation termination codons (PTCs) are recognized and eliminated by the nonsensemediated mRNA decay (NMD) pathway in eukaryotes. We employed a novel live-cell imaging approach to investigate the kinetics of mRNA synthesis and release at the transcription site of PTC-containing (PTC+) and PTC-free (PTC-) immunoglobulin-μ reporter genes. Fluorescence recovery after photobleaching (FRAP) and photoconversion analyses revealed that PTC+ transcripts are specifically retained at the transcription site. Remarkably, the retained PTC+ transcripts are mainly unspliced, and this RNA retention is dependent upon two important NMD factors, UPF1 and SMG6, since their depletion led to the release of the PTC+ transcripts. Finally, ChIP analysis showed a physical association of UPF1 and SMG6 with both the PTC+ and the PTC- reporter genes in vivo. Collectively, our data support a mechanism for regulation of PTC+ transcripts at the transcription site. Published by Cold Spring Harbor Laboratory Press.
Original language | English (US) |
---|---|
Pages (from-to) | 2094-2107 |
Number of pages | 14 |
Journal | RNA |
Volume | 17 |
Issue number | 12 |
DOIs | |
State | Published - Dec 2011 |
Keywords
- Live-cell imaging
- NMD
- Retention
- SMG6
- Splicing
- UPF1
ASJC Scopus subject areas
- Molecular Biology