4-thiouridine inhibits rRNA synthesis and causes a nucleolar stress response
| Autor: | Kaspar Burger, Michaela Rohrmoser, Dirk Eick, Bastian Mühl, Lukas Windhager, Caroline C. Friedel, Anita Gruber-Eber, Lars Dölken, Markus Kellner |
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| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: |
Nucleophosmin
Messenger RNA Staining and Labeling Nucleolus Cell Cycle Thiouridine RNA Ribosome biogenesis 4-thiouridine Ribosomal RNA rRNA processing p53 Nucleolar stress RNA labeling Cell Biology Biology Molecular biology Ribosome Mice RNA Ribosomal Stress Physiological Animals RNA Processing Post-Transcriptional RRNA processing Molecular Biology Ribosomes Cell Nucleolus Research Paper |
| Zdroj: | RNA Biol. 10, 1623-1630 (2013) |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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