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首页> 外文期刊>Molecular and Cellular Biology >Diazaborine Treatment of Yeast Cells Inhibits Maturation of the 60S Ribosomal Subunit
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Diazaborine Treatment of Yeast Cells Inhibits Maturation of the 60S Ribosomal Subunit

机译：Diazaborine处理酵母细胞可抑制60S核糖体亚基的成熟。

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Diazaborine treatment of yeast cells was shown previously to cause accumulation of aberrant, 3′-elongated mRNAs. Here we demonstrate that the drug inhibits maturation of rRNAs for the large ribosomal subunit. Pulse-chase analyses showed that the processing of the 27S pre-rRNA to consecutive species was blocked in the drug-treated wild-type strain. The steady-state level of the 7S pre-rRNA was clearly reduced after short-term treatment with the inhibitor. At the same time an increase of the 35S pre-rRNA was observed. Longer incubation with the inhibitor resulted in a decrease of the 27S precursor. Primer extension assays showed that an early step in 27S pre-rRNA processing is inhibited, which results in an accumulation of the 27SA2 pre-rRNA and a strong decrease of the 27SA3, 27SB1L, and 27SB1S precursors. The rRNA processing pattern observed after diazaborine treatment resembles that reported after depletion of the RNA binding protein Nop4p/Nop77p. This protein is essential for correct pre-27S rRNA processing. Using a green fluorescent protein-Nop4 fusion, we found that diazaborine treatment causes, within minutes, a rapid redistribution of the protein from the nucleolus to the periphery of the nucleus, which provides a possible explanation for the effect of diazaborine on rRNA processing.
机译：先前已显示用diazaborine处理酵母细胞会引起异常的3'延长mRNA积聚。在这里，我们证明该药物抑制大核糖体亚基的rRNA的成熟。脉冲追踪分析表明，在药物处理过的野生型菌株中，将27S pre-rRNA加工成连续物种的过程被阻止。抑制剂短期治疗后，7S pre-rRNA的稳态水平明显降低。同时观察到35S pre-rRNA的增加。与抑制剂长时间孵育会导致27S前体的减少。引物延伸分析表明，抑制了27S pre-rRNA加工的早期步骤，这导致27SA2 pre-rRNA的积累和27SA3、27SB1L和27SB1S前体的大量减少。在重氮唑嘌呤处理后观察到的rRNA加工模式类似于在耗尽RNA结合蛋白Nop4p / Nop77p后报道的rRNA处理模式。该蛋白对于正确的27S之前rRNA加工至关重要。使用绿色荧光蛋白-Nop4融合蛋白，我们发现重氮茶碱治疗可在几分钟内引起蛋白质从核仁快速重新分布到核的外围，这为重氮茶碱对rRNA加工的作用提供了可能的解释。

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