4-thiouridine inhibits rRNA synthesis and causes a nucleolar stress response
4-thiouridine inhibits rRNA synthesis and causes a nucleolar stress response
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DOI：
10.4161/rna.26214
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发表时间：
2013-10-01
期刊：
影响因子：
4.1
通讯作者：
Eick, Dirk
中科院分区：
文献类型：
--
作者：
Burger, Kaspar;Muehl, Bastian;Eick, Dirk
高浓度 (> 100 μM) 核糖核苷类似物 4-硫尿苷 (4sU) 广泛用于 RNA 分析方法，例如光活化核糖核苷增强交联和免疫沉淀 (PAR-CLIP) 和新生信使 (m)RNA 标记（4sU 标记）。在这里，我们证明低浓度 10 μM 的 4sU 标记可用于测量核糖体 (r)RNA 的生产和加工。然而，通常用于 mRNA 标记实验的 4sU (> 50 muM) 浓度升高会抑制 47S rRNA 的产生和加工。 rRNA 合成的抑制伴随着核仁核磷蛋白 (NPM1) 的核质易位、肿瘤抑制因子 p53 的诱导以及增殖的抑制。我们得出的结论是，4sU 对 RNA 的代谢标记会引发核仁应激反应，这可能会影响结果的解释。因此，功能性核糖体生物发生、核仁完整性和细胞周期应在 4sU 标记实验中解决。
High concentrations (> 100 mu M) 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 mu M can be used to measure production and processing of ribosomal (r)RNA. However, elevated concentrations of 4sU (> 50 mu M), 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.