核多聚 A 结合蛋白 Nab2 的纯化揭示了其与酵母转录组的关联及其信使核糖核蛋白核心结构。
Purification of nuclear poly(A)-binding protein Nab2 reveals association with the yeast transcriptome and a messenger ribonucleoprotein core structure.
机构信息
Biochemie Zentrum der Universität Heidelberg (BZH), Im Neuenheimer Feld 307, D-69120 Heidelberg, Germany.
出版信息
J Biol Chem. 2009 Dec 11;284(50):34911-7. doi: 10.1074/jbc.M109.062034. Epub 2009 Oct 19.
Abstract
Nascent mRNAs produced by transcription in the nucleus are subsequently processed and packaged into mRNA ribonucleoprotein particles (messenger ribonucleoproteins (mRNPs)) before export to the cytoplasm. Here, we have used the poly(A)-binding protein Nab2 to isolate mRNPs from yeast under conditions that preserve mRNA integrity. Upon Nab2-tandem affinity purification, several mRNA export factors were co-enriched (Yra1, Mex67, THO-TREX) that were present in mRNPs of different size and mRNA length. High-throughput sequencing of the co-precipitated RNAs indicated that Nab2 is associated with the bulk of yeast transcripts with no specificity for different mRNA classes. Electron microscopy revealed that many of the mRNPs have a characteristic elongated structure. Our data suggest that mRNPs, although associated with different mRNAs, have a unifying core structure.
摘要
在细胞核中转录产生的新生 mRNA 随后经过加工并包装成 mRNA 核糖核蛋白颗粒（信使核糖核蛋白（mRNPs）），然后输出到细胞质。在这里，我们使用多聚（A）结合蛋白 Nab2 在保持 mRNA 完整性的条件下从酵母中分离 mRNPs。在 Nab2 串联亲和纯化后，几种 mRNA 输出因子（Yra1、Mex67、THO-TREX）被共富集，这些因子存在于不同大小和 mRNA 长度的 mRNPs 中。共沉淀 RNA 的高通量测序表明，Nab2 与大量酵母转录物相关，而对不同的 mRNA 类别没有特异性。电子显微镜显示，许多 mRNPs 具有特征性的细长结构。我们的数据表明，mRNPs 尽管与不同的 mRNA 相关，但具有统一的核心结构。
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