Lafontaine D L, Tollervey D
Institute of Cell and Molecular Biology, The University of Edinburgh, Scotland.
RNA. 1999 Mar;5(3):455-67. doi: 10.1017/s135583829998192x.
Eukaryotic nucleoli contain a large family of box C+D small nucleolar RNA (snoRNA) species, all of which are associated with a common protein Nop1p/fibrillarin. Nop58p was identified in a screen for synthetic lethality with Nop1p and shown to be an essential nucleolar protein. Here we report that a Protein A-tagged version of Nop58p coprecipitates all tested box C+D snoRNAs and that genetic depletion of Nop58p leads to the loss of all tested box C+D snoRNAs. The box H+ACA class of snoRNAs are not coprecipitated with Nop58p, and are not codepleted. The yeast box C+D snoRNAs include two species, U3 and U14, that are required for the early cleavages in pre-rRNA processing. Consistent with this, Nop58p depletion leads to a strong inhibition of pre-rRNA processing and 18S rRNA synthesis. Unexpectedly, depletion of Nop58p leads to the accumulation of 3' extended forms of U3 and U24, showing that the protein is also involved in snoRNA synthesis. Nop58p is the second common component of the box C+D snoRNPs to be identified and the first to be shown to be required for the stability and for the synthesis of these snoRNAs.
真核生物的核仁包含一个由盒C+D小核仁RNA(snoRNA)种类组成的大家族,所有这些都与一种共同的蛋白质Nop1p/纤维原蛋白相关联。Nop58p是在与Nop1p的合成致死筛选中被鉴定出来的,并被证明是一种必需的核仁蛋白。在此我们报告,一个带有蛋白A标签的Nop58p版本能共沉淀所有测试的盒C+D snoRNA,并且Nop58p的基因缺失会导致所有测试的盒C+D snoRNA的丢失。盒H+ACA类的snoRNA不会与Nop58p共沉淀,也不会被共同耗尽。酵母盒C+D snoRNA包括两种,U3和U14,它们是前体rRNA加工早期切割所必需的。与此一致的是,Nop58p的缺失会导致前体rRNA加工和18S rRNA合成受到强烈抑制。出乎意料的是,Nop58p的缺失会导致U3和U24的3'延伸形式的积累,表明该蛋白也参与snoRNA的合成。Nop58p是第二个被鉴定出来的盒C+D snoRNP的共同成分,也是第一个被证明对这些snoRNA的稳定性和合成是必需的成分。