The essential WD-repeat protein Rsa4p is required for rRNA processing and intra-nuclear transport of 60S ribosomal subunits.
The essential WD-repeat protein Rsa4p is required for rRNA processing and intra-nuclear transport of 60S ribosomal subunits.
复制标题
rRNA 加工和 60S 核糖体亚基的核内运输需要必需的 WD 重复蛋白 Rsa4p。
DOI：
10.1093/nar/gki887
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发表时间：
2005
影响因子：
14.9
通讯作者：
Remacha M
中科院分区：
文献类型：
--
作者：
de la Cruz J;Sanz-Martínez E;Remacha M
我们报道了一种新的因子，Rsa4p(Ycr072cp)，它是合成60S核糖体亚基所必需的。Rsa4p是一种保守的WD重复蛋白，似乎定位于核仁中。在体内，Rsa4p的耗尽会导致60S核糖体亚基的缺失和半聚体多聚体的出现。对前-rRNA和成熟rRNA的Northern杂交和引物延伸分析表明，Rsa4p的缺失导致27S、25.5S和7S前rRNA的积累，导致成熟的25S和5.8S rRNA减少。对Pre-rRNA加工的脉冲追逐分析表明，至少这是由于27S Pre-rRNA中间产物到成熟的25S rRNA的强烈延迟。此外，Rsa4p的耗尽抑制了60年代前核糖体颗粒从核仁到核质的释放，这主要是从大亚基Rpl25-EGFP报告结构的核仁积累来判断的。我们认为Rsa4p早期与60s前的核糖体颗粒结合，并为rRNA前体的正确处理和60s核糖体亚基的核仁释放提供了一个相互作用的平台。
We report the characterization of a novel factor, Rsa4p (Ycr072cp), which is essential for the synthesis of 60S ribosomal subunits. Rsa4p is a conserved WD-repeat protein that seems to localize in the nucleolus. In vivo depletion of Rsa4p results in a deficit of 60S ribosomal subunits and the appearance of half-mer polysomes. Northern hybridization and primer extension analyses of pre-rRNA and mature rRNAs show that depletion of Rsa4p leads to the accumulation of the 27S, 25.5S and 7S pre-rRNAs, resulting in a reduction of the mature 25S and 5.8S rRNAs. Pulse–chase analyses of pre-rRNA processing reveal that, at least, this is due to a strong delay in the maturation of 27S pre-rRNA intermediates to mature 25S rRNA. Furthermore, depletion of Rsa4p inhibited the release of the pre-60S ribosomal particles from the nucleolus to the nucleoplasm, as judged by the predominantly nucleolar accumulation of the large subunit Rpl25-eGFP reporter construct. We propose that Rsa4p associates early with pre-60S ribosomal particles and provides a platform of interaction for correct processing of rRNA precursors and nucleolar release of 60S ribosomal subunits.
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