Composition and functional characterization of yeast 66S ribosome assembly intermediates
Composition and functional characterization of yeast 66S ribosome assembly intermediates
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DOI：
10.1016/s1097-2765(01)00344-6
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发表时间：
2001-09-01
期刊：
影响因子：
16
通讯作者：
Woolford, JL
中科院分区：
文献类型：
--
作者：
Harnpicharnchai, P;Jakovljevic, J;Woolford, JL
核糖体组装的途径和完整的因素集合尚不清楚。为了解决这些问题，我们亲和纯化的酵母前核糖体颗粒含有核仁蛋白Nop7p和开发的手段来分离它们的组件。Nop7p主要与含有27SB或25.5S加7S前rRNA的66S前核糖体相关。通过质谱鉴定的共纯化蛋白包括核糖体蛋白、先前与60S核糖体生物发生有关的非核糖体蛋白和未知参与核糖体产生的蛋白。对这些蛋白质中的8种突变株的分析表明，它们确实参与了核糖体生物合成。这些结果表明，蛋白质组学方法与遗传工具相结合，提供了强有力的手段来纯化和表征核糖体组装中间体。
The pathway and complete collection of factors that orchestrate ribosome assembly are not clear. To address these problems, we affinity purified yeast preribosomal particles containing the nucleolar protein Nop7p and developed means to separate their components. Nop7p is associated primarily with 66S preribosomes containing either 27SB or 25.5S plus 7S pre-rRNAs. Copurifying proteins identified by mass spectrometry include ribosomal proteins, nonribosomal proteins previously implicated in 60S ribosome biogenesis, and proteins not known to be involved in ribosome production. Analysis of strains mutant for eight of these proteins not previously implicated in ribosome biogenesis showed that they do participate in this pathway. These results demonstrate that proteomic approaches in concert with genetic tools provide powerful means to purify and characterize ribosome assembly intermediates.