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首页> 外文期刊>EMBO Journal >Architecture and assembly of mammalian H/ACA small nucleolar and telomerase ribonucleoproteins
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Architecture and assembly of mammalian H/ACA small nucleolar and telomerase ribonucleoproteins

机译：哺乳动物的体系结构和装配H / ACA小核仁的端粒酶和核糖核蛋白

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摘要

Mammalian H/ACA small nucleolar RNAs and telomerase RNA share common sequence and secondary structure motifs that form ribonucleoprotein particles (RNPs) with the same four core proteins, NAP57 (also dyskerin or in yeast Cbf5p), GAR1, NHP2, and NOP10. The assembly and molecular interactions of the components of H/ACA RNPs are unknown. Using in vitro transcription/translation in combination with immunoprecipitation of core proteins, UV-crosslinking, and electrophoretic mobility shift assays, we demonstrate the following. NOP10 associates with NAP57 as a prerequisite for NHP2 binding. Although NHP2 on its own binds RNA nonspecifically, this NAP57 NOP10-NHP2 core trimer specifically recognizes H/ACA RNAs. GAR1 associates independently with NAP57 near the pseudouridylase core of mature H/ACA RNPs. In contrast to other RNPs whose assembly is initiated by protein RNA interactions, the four H/ACA core proteins form a protein-only particle that associates with H/ACA RNAs. Nonetheless, functional H/ACA snoRNPs assembled in cytosolic extracts are stable and do not exchange their RNA components, suggesting that new particle formation requires de novo synthesis.
机译：哺乳动物的H / ACA小核仁的rna端粒酶RNA共同序列和二级结构形成的图案核糖核蛋白粒子(rnp)相同四个核心蛋白质,NAP57(也dyskerin或酵母Cbf5p)、GAR1 NHP2, NOP10。和分子的交互组件H / ACA rnp是未知的。转录/翻译结合免疫沉淀反应的核心蛋白质,UV-crosslinking,电泳迁移率转变化验,我们证明如下。同事与NAP57 NHP2的先决条件绑定。是非,这NAP57 NOP10-NHP2核心三聚物特别认识到H / ACA rna。将独立与NAP57附近pseudouridylase H / ACA rnp成熟的核心。相比其他rnp的组装由蛋白质的RNA相互作用,四个H / ACA核心蛋白质形成protein-only粒子同事与H / ACA rna。功能性H / ACA snoRNPs聚集在胞质提取是稳定的,不交换他们的RNA组件,这表明新粒子形成需要从头合成。

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