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

Abstract:

: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.

journal_name

RNA Biol

journal_title

RNA biology

authors

Burger K,Mühl B,Kellner M,Rohrmoser M,Gruber-Eber A,Windhager L,Friedel CC,Dölken L,Eick D

doi

10.4161/rna.26214

subject

Has Abstract

pub_date

2013-10-01 00:00:00

pages

1623-30

issue

10

eissn

1547-6286

issn

1555-8584

pii

26214

journal_volume

10

pub_type

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