Institute of Biological Chemistry, Washington State University, Pullman, Washington 99164-6340, USA.
J Proteome Res. 2009 Oct;8(10):4641-53. doi: 10.1021/pr900537p.
In eukaryotes, RNA binding proteins (RBPs) play an integral role not only in RNA processing within the nucleus, but also in the cytoplasmic events of RNA transport, localization, translation, storage and degradation. While many studies have been done, relatively little is known about RBPs in plants. As part of our continuing efforts to understand cytoplasmic gene expression events in developing rice seed (Oryza sativa L.), a proteomics approach was used to identify cytoplasmic-localized, cytoskeletal-associated RBPs. The nucleic acid binding fraction from a cytoskeletal-enriched rice seed extract was isolated by Poly(U)-Sepharose affinity chromatography and analyzed using 2D gel electrophoresis. Analysis of 162 excised protein spots using mass spectrometry led to the identification of 148 distinct proteins, in addition to the highly abundant globulin and glutelin seed storage proteins. Identified proteins include those involved in RNA processing, translation, protein modification, cell signaling, and metabolism, as well as a number of hypothetical proteins. Proteins of particular interest with roles in RNA metabolism are discussed. These results have been deposited within the Rice RNA Binding Protein Database as part of an integrated study of plant cytoskeletal-associated RBPs using developing rice seed as a model.
在真核生物中，RNA 结合蛋白 (RBP) 不仅在核内的 RNA 处理中发挥着重要作用，而且在 RNA 运输、定位、翻译、储存和降解的细胞质事件中也发挥着重要作用。尽管已经进行了许多研究，但对植物中的 RBP 相对知之甚少。作为我们持续努力理解发育中的水稻种子（Oryza sativa L.）细胞质基因表达事件的一部分，采用蛋白质组学方法来鉴定细胞质定位的、与细胞骨架相关的 RBP。通过 Poly(U)-Sepharose 亲和层析从富含细胞骨架的水稻种子提取物中分离出核酸结合部分，并使用 2D 凝胶电泳进行分析。使用质谱法分析 162 个切除的蛋白斑点，鉴定出 148 种不同的蛋白质，除了高度丰富的球蛋白和谷蛋白种子储存蛋白外。鉴定出的蛋白质包括参与 RNA 处理、翻译、蛋白质修饰、细胞信号转导和代谢的蛋白质，以及许多假设蛋白。讨论了在 RNA 代谢中具有特定作用的感兴趣的蛋白质。这些结果已作为植物细胞骨架相关 RBP 综合研究的一部分，存储在 Rice RNA Binding Protein Database 中，该研究以发育中的水稻种子为模型。
