Stepwise RNP assembly at the site of H/ACA RNA transcription in human cells.

Stepwise RNP assembly at the site of H/ACA RNA transcription in human cells.
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DOI:
10.1083/jcb.200601105
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发表时间:
2006-04-24
期刊:
The Journal of cell biology
影响因子:
--
通讯作者:
Meier UT
Meier UT
中科院分区:
其他
文献类型:
--
作者:
Darzacq X;Kittur N;Roy S;Shav-Tal Y;Singer RH;Meier UT

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哺乳动物的H/ACA核糖核蛋白(RNP)对核糖体生物发生、前信使RNA剪接以及端粒维持至关重要。这些RNP由四种核心蛋白和一种RNA组成,但它们如何组装尚不清楚。通过对H/ACA RNA转录位点的研究,我们解析了它们在单细胞中的生物发生过程,并阐明了非核心蛋白NAF1在此过程中的作用。NAF1以及除GAR1之外的所有核心蛋白都被招募到转录位点。NAF1与核心蛋白之一NAP57结合,并在细胞核和细胞质之间穿梭。这两种蛋白对稳定的H/ACA RNA积累都至关重要。NAF1和GAR1竞争性地结合NAP57,这表明存在一种顺序性的相互作用。我们的分析表明,NAF1结合NAP57并将其护送到新生的H/ACA RNA处,然后GAR1取代NAF1,从而在卡哈尔体和核仁中产生成熟的H/ACA RNP。
Mammalian H/ACA RNPs are essential for ribosome biogenesis, premessenger RNA splicing, and telomere maintenance. These RNPs consist of four core proteins and one RNA, but it is not known how they assemble. By interrogating the site of H/ACA RNA transcription, we dissected their biogenesis in single cells and delineated the role of the non-core protein NAF1 in the process. NAF1 and all of the core proteins except GAR1 are recruited to the site of transcription. NAF1 binds one of the core proteins, NAP57, and shuttles between nucleus and cytoplasm. Both proteins are essential for stable H/ACA RNA accumulation. NAF1 and GAR1 bind NAP57 competitively, suggesting a sequential interaction. Our analyses indicate that NAF1 binds NAP57 and escorts it to the nascent H/ACA RNA and that GAR1 then replaces NAF1 to yield mature H/ACA RNPs in Cajal bodies and nucleoli.
NOPP140充当核仁与盘绕体之间的分子联系。
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