Potent, Reversible, and Specific Chemical Inhibitors of Eukaryotic Ribosome Biogenesis.
Potent, Reversible, and Specific Chemical Inhibitors of Eukaryotic Ribosome Biogenesis.
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DOI：
10.1016/j.cell.2016.08.070
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发表时间：
2016-10-06
期刊：
影响因子：
64.5
通讯作者：
Kapoor, Tarun M.
中科院分区：
文献类型：
--
作者：
Kawashima, Shigehiro A.;Chen, Zhen;Aoi, Yuki;Patgiri, Anupam;Kobayashi, Yuki;Nurse, Paul;Kapoor, Tarun M.
所有的细胞蛋白质都是由核糖体合成的，真核生物的核糖体合成是一个复杂的多步骤过程，在几分钟内完成。有几种核糖体功能的化学抑制剂可用作工具或药物。相比之下，我们缺乏有效的验证化学探针来分析真核生物核糖体组装的动力学。在这里，我们结合联合收割机化学和遗传学的方法来发现ribozinoindoles（或RBIN），有效的和可逆的三嗪吲哚为基础的真核生物核糖体生物合成的抑制剂。对Rbin敏感性和抗性赋予裂殖酵母突变的分析，沿着重组蛋白的生化测定，提供了Rbin的生理靶点是Midasin的证据，Midasin是一种必需的540-kDa AAA+（与多种细胞活性相关的ATP酶）蛋白。使用RBIN来急性抑制或激活Midasin功能，在与细胞敏感或细胞耐药细胞的平行实验中，我们揭示了Midasin在组装Nsa 1颗粒（60 S亚基的核仁前体）中的作用。总之，我们的研究结果表明，RBIN是真核生物核糖体组装的强大探针。Ribozinoindoles是真核生物核糖体组装的有效化学抑制剂Mdn 1的六个ATP酶位点中的四个的活性可能是细胞生长所需的Ribozinoindoles抑制重组全长Mdn 1的ATP酶活性体外组装Nsa 1颗粒，60 S亚基的前体，依赖于Mdn 1通过化学合成致死筛选发现了真核生物核糖体生物合成的选择性有效抑制剂。
All cellular proteins are synthesized by ribosomes, whose biogenesis in eukaryotes is a complex multi-step process completed within minutes. Several chemical inhibitors of ribosome function are available and used as tools or drugs. By contrast, we lack potent validated chemical probes to analyze the dynamics of eukaryotic ribosome assembly. Here, we combine chemical and genetic approaches to discover ribozinoindoles (or Rbins), potent and reversible triazinoindole-based inhibitors of eukaryotic ribosome biogenesis. Analyses of Rbin sensitivity and resistance conferring mutations in fission yeast, along with biochemical assays with recombinant proteins, provide evidence that Rbins’ physiological target is Midasin, an essential ∼540-kDa AAA+ (ATPases associated with diverse cellular activities) protein. Using Rbins to acutely inhibit or activate Midasin function, in parallel experiments with inhibitor-sensitive or inhibitor-resistant cells, we uncover Midasin’s role in assembling Nsa1 particles, nucleolar precursors of the 60S subunit. Together, our findings demonstrate that Rbins are powerful probes for eukaryotic ribosome assembly. Ribozinoindoles are potent chemical inhibitors of eukaryotic ribosome assembly Activity of four of Mdn1’s six ATPase sites is likely needed for cell growth Ribozinoindoles inhibit recombinant full-length Mdn1’s ATPase activity in vitro Assembly of Nsa1 particles, precursors of the 60S subunit, depends on Mdn1 Selective potent inhibitors of eukaryotic ribosome biogenesis are found through a chemical synthetic lethal screen.
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影响因子：
4.8
作者：
Galani, K;Nissan, TA;Hurt, E
通讯作者：
Hurt, E
DOI：
10.1016/j.bbamcr.2011.06.017
发表时间：
2012-01
期刊：
Biochimica et biophysica acta
影响因子：
--
作者：
Kressler D;Hurt E;Bergler H;Bassler J
通讯作者：
Bassler J
影响因子：
11.4
作者：
Funabiki, H;Kumada, K;Yanagida, M
通讯作者：
Yanagida, M
影响因子：
19
作者：
Patel, S;Latterich, M
通讯作者：
Latterich, M
影响因子：
4
作者：
Thomson, Emma;Ferreira-Cerca, Sebastien;Hurt, Ed
通讯作者：
Hurt, Ed