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首页> 美国卫生研究院文献>The Journal of Biophysical and Biochemical Cytology >The Telomerase/Vault-Associated Protein Tep1 Is Required for Vault RNA Stability and Its Association with the Vault Particle
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The Telomerase/Vault-Associated Protein Tep1 Is Required for Vault RNA Stability and Its Association with the Vault Particle

机译：保管库RNA稳定性及其与保管库颗粒的关联需要端粒酶/保管库相关蛋白Tep1

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摘要

Vaults and telomerase are ribonucleoprotein (RNP) particles that share a common protein subunit, TEP1. Although its role in either complex has not yet been defined, TEP1 has been shown to interact with the mouse telomerase RNA and with several of the human vault RNAs in a yeast three-hybrid assay. An mTep1 −/− mouse was previously generated which resulted in no apparent change in telomere length or telomerase activity in six generations of mTep1-deficient mice. Here we show that the levels of the telomerase RNA and its association with the telomerase RNP are also unaffected in mTep1 − /− mice. Although vaults purified from the livers of mTep1−/− mice appear structurally intact by both negative stain and cryoelectron microscopy, three-dimensional reconstruction of the mTep1−/− vault revealed less density in the cap than previously observed for the intact rat vault. Furthermore, the absence of TEP1 completely disrupted the stable association of the vault RNA with the purified vault particle and also resulted in a decrease in the levels and stability of the vault RNA. Therefore, we have uncovered a novel role for TEP1 in vivo as an integral vault protein important for the stabilization and recruitment of the vault RNA to the vault particle.
机译：穹顶和端粒酶是核糖核蛋白（RNP）颗粒，它们共有一个共同的蛋白质亚基TEP1。尽管尚未确定其在两种复合物中的作用，但在酵母三杂交试验中，TEP1已显示与小鼠端粒酶RNA以及与某些人类穹顶RNA相互作用。先前生成了mTep1 -/-小鼠，该小鼠在6代mTep1缺陷小鼠中没有导致端粒长度或端粒酶活性的明显变化。在这里我们显示端粒酶RNA的水平及其与端粒酶RNP的关联在mTep1 - / -小鼠中也不受影响。尽管从mTep1 -/-小鼠肝脏中纯化的穹顶在阴性染色和冷冻电子显微镜下均在结构上完整无缺，但mTep1 -/-穹顶的三维重建显示较少帽中的密度比完整的大鼠穹顶先前观察到的密度高。此外，TEP1的缺失完全破坏了穹顶RNA与纯化的穹顶颗粒的稳定结合，并且还导致了穹顶RNA的水平和稳定性下降。因此，我们发现了TEP1在体内作为一种重要的整体金库蛋白质，对于稳定金库RNA和将金库RNA募集到金库粒子具有重要作用。

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