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Open Access Green as soon as Postprint is submitted to ZB.
4-thiouridine inhibits rRNA synthesis and causes a nucleolar stress response.
RNA Biol. 10, 1623-1630 (2013)
High concentrations (> 100 µM) of the ribonucleoside analog 4-thiouridine (4sU) is widely used in methods for RNA analysis like photoactivatable-ribonucleoside-enhanced crosslinking and immunoprecipitation (PAR-CLIP) and nascent messenger (m)RNA labeling (4sU-tagging). Here, we show that 4sU-tagging at low concentrations ≤ 10 µM can be used to measure production and processing of ribosomal (r)RNA. However, elevated concentrations of 4sU (> 50 µM), which are usually used for mRNA labeling experiments, inhibit production and processing of 47S rRNA. The inhibition of rRNA synthesis is accompanied by nucleoplasmic translocation of nucleolar nucleophosmin (NPM1), induction of the tumor suppressor p53, and inhibition of proliferation. We conclude that metabolic labeling of RNA by 4sU triggers a nucleolar stress response, which might influence the interpretation of results. Therefore, functional ribosome biogenesis, nucleolar integrity, and cell cycle should be addressed in 4sU labeling experiments.
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Publication type
Article: Journal article
Document type
Scientific Article
Keywords
4-thiouridine; Ribosomal RNA; rRNA processing; p53; Nucleolar stress; RNA labeling; Nucleophosmin; Microarray Analysis ; Cell-proliferation ; P53 Activation ; Protein L11 ; Degradation ; Biogenesis ; Pseudouridylation ; 5-fluorouracil ; Resolution ; Binding
ISSN (print) / ISBN
1547-6286
e-ISSN
1555-8584
Journal
RNA Biology
Quellenangaben
Volume: 10,
Issue: 10,
Pages: 1623-1630
Publisher
Landes Bioscience
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Reviewing status
Peer reviewed