Gosai Sager J, Foley Shawn W, Wang Dongxue, Silverman Ian M, Selamoglu Nur, Nelson Andrew D L, Beilstein Mark A, Daldal Fevzi, Deal Roger B, Gregory Brian D
Department of Biology, University of Pennsylvania, Philadelphia, PA 19104, USA.
Department of Biology, University of Pennsylvania, Philadelphia, PA 19104, USA; Cell and Molecular Biology Graduate Group, University of Pennsylvania, Philadelphia, PA 19104, USA.
Mol Cell. 2015 Jan 22;57(2):376-88. doi: 10.1016/j.molcel.2014.12.004. Epub 2014 Dec 31.
Posttranscriptional regulation in eukaryotes requires cis- and trans-acting features and factors including RNA secondary structure and RNA-binding proteins (RBPs). However, a comprehensive view of the structural and RBP interaction landscape of nuclear RNAs has yet to be compiled for any organism. Here, we use our ribonuclease-mediated structure and RBP-binding site mapping approaches to globally profile these features in Arabidopsis seedling nuclei in vivo. We reveal anticorrelated patterns of secondary structure and RBP binding throughout nuclear mRNAs that demarcate sites of alternative splicing and polyadenylation. We also uncover a collection of protein-bound sequence motifs, and identify their structural contexts, co-occurrences in transcripts encoding functionally related proteins, and interactions with putative RBPs. Finally, using these motifs, we find that the chloroplast RBP CP29A also interacts with nuclear mRNAs. In total, we provide a simultaneous view of the RNA secondary structure and RBP interaction landscapes in a eukaryotic nucleus.
真核生物中的转录后调控需要顺式和反式作用特征及因子,包括RNA二级结构和RNA结合蛋白(RBPs)。然而,尚未针对任何生物体汇编出核RNA的结构和RBP相互作用图谱的全面视图。在此,我们使用核糖核酸酶介导的结构和RBP结合位点映射方法,在体内对拟南芥幼苗细胞核中的这些特征进行全局分析。我们揭示了整个核mRNA中二级结构和RBP结合的反相关模式,这些模式划分了可变剪接和多聚腺苷酸化的位点。我们还发现了一组蛋白质结合序列基序,并确定了它们的结构背景、在编码功能相关蛋白质的转录本中的共现情况以及与假定RBP的相互作用。最后,利用这些基序,我们发现叶绿体RBP CP29A也与核mRNA相互作用。总之,我们提供了真核细胞核中RNA二级结构和RBP相互作用图谱的同时视图。