Cotranscriptional effect of a premature termination codon revealed by live-cell imaging

被引：39

作者：

de Turris, Valeria ^{[1
]}

Nicholson, Pamela ^{[2
]}

Orozco, Rodolfo Zamudio ^{[2
]}

Singer, Robert H. ^{[1
]}

Muehlemann, Oliver ^{[2
]}

机构：

[1] Albert Einstein Coll Med, Bronx, NY 10461 USA

[2] Univ Bern, Dept Chem & Biochem, CH-3012 Bern, Switzerland

来源：

RNA | 2011年 / 17卷 / 12期

基金：

欧洲研究理事会; 瑞士国家科学基金会;

关键词：

live-cell imaging; NMD; retention; splicing; UPF1; SMG6; NONSENSE-MEDIATED DECAY; MESSENGER-RNA DECAY; EXON-JUNCTION COMPLEX; OPEN READING FRAME; LIGHT-CHAIN GENES; MAMMALIAN-CELLS; QUALITY-CONTROL; PROTEIN-SYNTHESIS; HALF-LIFE; TRANSLATION;

D O I：

10.1261/rna.02918111

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Aberrant mRNAs with premature translation termination codons (PTCs) are recognized and eliminated by the nonsense-mediated 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-mu 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.

引用

页码：2094 / 2107

页数：14

共 64 条

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