2013/09/11 by Kaspar Burger, Bastian Mühl, Markus Kellner +6 · 140 citations
Biochemistry, Genetics and Molecular Biology · #RNA modifications and cancer #RNA and protein synthesis mechanisms #RNA Research and Splicing #Ribosome biogenesis #Biology #Nucleophosmin #Ribosomal RNA #Nucleolus #RNA #Ribonucleoside #Ribosome #Protein biosynthesis #Cell biology #Molecular biology #Messenger RNA #Biogenesis #Biochemistry #Gene #Cytoplasm
paper · pdf · doi:10.4161/rna.26214
published in RNA Biology 10(10), 1623-1630 (Taylor & Francis)
openalex publication_date 2013/09/11 · openalex created_date 2025/10/10 · openalex updated_date 2026/08/05
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.