Department of Microbiology and Molecular Genetics, UMDNJ-NJ Medical School, Newark, New Jersey 07103, USA.
RNA. 2012 Nov;18(11):1968-83. doi: 10.1261/rna.033829.112. Epub 2012 Sep 10.
RNA-binding proteins that target mRNA coding regions are emerging as regulators of post-transcriptional processes in eukaryotes. Here we describe a newly identified RNA-binding protein, RBP42, which targets the coding region of mRNAs in the insect form of the African trypanosome, Trypanosoma brucei. RBP42 is an essential protein and associates with polysome-bound mRNAs in the cytoplasm. A global survey of RBP42-bound mRNAs was performed by applying HITS-CLIP technology, which captures protein-RNA interactions in vivo using UV light. Specific RBP42-mRNA interactions, as well as mRNA interactions with a known RNA-binding protein, were purified using specific antibodies. Target RNA sequences were identified and quantified using high-throughput RNA sequencing. Analysis revealed that RBP42 bound mainly within the coding region of mRNAs that encode proteins involved in cellular energy metabolism. Although the mechanism of RBP42's function is unclear at present, we speculate that RBP42 plays a critical role in modulating T. brucei energy metabolism.
在真核生物中,靶向 mRNA 编码区的 RNA 结合蛋白正在成为转录后过程的调控因子。在这里,我们描述了一种新鉴定的 RNA 结合蛋白 RBP42,它靶向非洲锥虫昆虫形式的 mRNA 的编码区。RBP42 是一种必需蛋白,与细胞质中多核糖体结合的 mRNA 相关联。通过应用 HITS-CLIP 技术对 RBP42 结合的 mRNA 进行了全面调查,该技术使用紫外线在体内捕获蛋白-RNA 相互作用。使用特异性抗体纯化特定的 RBP42-mRNA 相互作用以及与已知 RNA 结合蛋白的 mRNA 相互作用。使用高通量 RNA 测序鉴定和定量靶 RNA 序列。分析表明,RBP42 主要结合编码参与细胞能量代谢的蛋白质的 mRNA 的编码区。虽然目前 RBP42 功能的机制尚不清楚,但我们推测 RBP42 在调节 T. brucei 能量代谢中发挥关键作用。