Has1 regulates consecutive maturation and processing steps for assembly of 60S ribosomal subunits.

Has1 regulates consecutive maturation and processing steps for assembly of 60S ribosomal subunits.
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HAS1调节连续的成熟和处理步骤,用于组装60S核糖体亚基。

DOI:
10.1093/nar/gkt545
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发表时间:
2013-09
影响因子:
14.9
通讯作者:
Woolford JL Jr
Woolford JL Jr
中科院分区:
生物学2区
文献类型:
--
作者:
Dembowski JA;Kuo B;Woolford JL Jr

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酿酒酵母核糖体的生物发生需要∼200组装因子。与这些因子中的大多数缺失相关的前核糖体RNA(RRNA)加工缺陷已经被表征。然而,组装因素如何驱动核糖核蛋白邻域的构建,以及结构重排如何与rRNA前加工相耦合,目前尚不清楚。在这里,我们揭示了Has1 DEAD-box RNA解旋酶在构建60S核糖体亚基的连续前rRNA加工和成熟步骤中的ATP依赖和ATP依赖的作用。Has1以一种不依赖于ATP的方式与60年代前的核糖体结合。Has1结合触发27SA3前-rRNA的核酸外切,产生5.8S rRNA的5‘端,并驱动5.8S/25S rRNA结构域I的核糖体蛋白L17的掺入。Has1的ATP依赖活性促进了多肽出口隧道周围额外结构域I核糖体蛋白的稳定结合,这是27SB pre-rRNA下游加工所必需的。此外,在缺乏Has1的情况下,异常的27S前rRNAs被定位为不可逆转的翻转。因此,我们的数据支持一种模型,在该模型中,Has1帮助建立结构域I结构,以防止前rRNA周转,并将结构域I折叠与连续的前rRNA处理步骤相结合。
Ribosome biogenesis requires ∼200 assembly factors in Saccharomyces cerevisiae. The pre-ribosomal RNA (rRNA) processing defects associated with depletion of most of these factors have been characterized. However, how assembly factors drive the construction of ribonucleoprotein neighborhoods and how structural rearrangements are coupled to pre-rRNA processing are not understood. Here, we reveal ATP-independent and ATP-dependent roles of the Has1 DEAD-box RNA helicase in consecutive pre-rRNA processing and maturation steps for construction of 60S ribosomal subunits. Has1 associates with pre-60S ribosomes in an ATP-independent manner. Has1 binding triggers exonucleolytic trimming of 27SA3 pre-rRNA to generate the 5′ end of 5.8S rRNA and drives incorporation of ribosomal protein L17 with domain I of 5.8S/25S rRNA. ATP-dependent activity of Has1 promotes stable association of additional domain I ribosomal proteins that surround the polypeptide exit tunnel, which are required for downstream processing of 27SB pre-rRNA. Furthermore, in the absence of Has1, aberrant 27S pre-rRNAs are targeted for irreversible turnover. Thus, our data support a model in which Has1 helps to establish domain I architecture to prevent pre-rRNA turnover and couples domain I folding with consecutive pre-rRNA processing steps.
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