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Burger, K. ; Mühl, B. ; Kellner, M. ; Rohrmoser, M. ; Gruber-Eber, A. ; Windhager, L.* ; Friedel, C.C.* ; Dölken, L.* ; Eick, D.

4-thiouridine inhibits rRNA synthesis and causes a nucleolar stress response.

RNA Biol. 10, 1623-1630 (2013)
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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
Language english
Publication Year 2013
HGF-reported in Year 2013
ISSN (print) / ISBN 1547-6286
e-ISSN 1555-8584
Journal RNA Biology
Quellenangaben Volume: 10, Issue: 10, Pages: 1623-1630 Article Number: , Supplement: ,
Publisher Landes Bioscience
Reviewing status Peer reviewed
Institute(s) Institute of Clinical Molecular Biology and Tumor Genetics (K.MOLBI)
POF-Topic(s) 30504 - Mechanisms of Genetic and Environmental Influences on Health and Disease
Research field(s) Immune Response and Infection
PSP Element(s) G-501490-001
DOI 10.4161/rna.26214
WOS ID WOS:000327369000009
Scopus ID 84886044727
PubMed ID 24025460
Erfassungsdatum 2013-10-16
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