Tompkins V S, Valverde D P, Moss W N
Roy J. Carver Department of Biochemistry, Biophysics, and Molecular Biology, Iowa State University, 2437 Pammel Drive, Ames, IA, 50011, USA.
Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, CT, 06536, USA.
BMC Res Notes. 2018 Feb 20;11(1):139. doi: 10.1186/s13104-018-3250-8.
The function of Epstein-Barr virus (EBV) stable intronic sequence (sis)RNAs, non-coding RNAs transcribed from a region required for EBV-mediated cellular transformation, remain unknown. To better understand the function of ebv-sisRNA-1 and ebv-sisRNA-2 from the internal repeat (IR)1 region of EBV, we used a combination of bioinformatics and biochemistry to identify associated RNA binding proteins. The findings reported here are part of ongoing studies to determine the functions of non-coding RNAs from the IR1 region of EBV.
Human regulatory proteins HNRNPA1 (heterogeneous nuclear ribonucleoprotein A1), HNRNPC, HNRNPL, HuR (human antigen R), and protein LIN28A (lin-28 homolog A) were predicted to bind ebv-sisRNA-1 and/or ebv-sisRNA-2; FUS (fused in sarcoma) was predicted to associate with ebv-sisRNA-2. Protein interactions were validated using a combination of RNA immunoprecipitation and biotin pulldown assays. Both sisRNAs also precipitated with HNRNPD and NONO (non-POU domain-containing octamer-binding protein). Interestingly, each of these interacting proteins also precipitated non-spliced non-coding RNA sequences transcribed from the IR1 region. Our findings suggest interesting roles for sisRNAs (through their interactions with regulatory proteins) and provide further evidence for the existence of non-spliced stable non-coding RNAs.
爱泼斯坦-巴尔病毒（EBV）稳定内含子序列（sis）RNAs是从EBV介导的细胞转化所需区域转录而来的非编码RNAs，其功能尚不清楚。为了更好地了解来自EBV内部重复序列（IR）1区域的ebv-sisRNA-1和ebv-sisRNA-2的功能，我们结合生物信息学和生物化学方法来鉴定相关的RNA结合蛋白。本文报道的研究结果是确定来自EBV的IR1区域非编码RNA功能的正在进行的研究的一部分。
预测人类调节蛋白HNRNPA1（异质性核核糖核蛋白A1）、HNRNPC、HNRNPL、HuR（人类抗原R）和蛋白LIN28A（lin-28同源物A）与ebv-sisRNA-1和/或ebv-sisRNA-2结合；预测FUS（肉瘤融合蛋白）与ebv-sisRNA-2相关。使用RNA免疫沉淀和生物素下拉试验相结合的方法验证了蛋白质相互作用。两种sisRNAs也与HNRNPD和NONO（含非POU结构域的八聚体结合蛋白）一起沉淀。有趣的是，这些相互作用的蛋白中的每一种也沉淀了从IR1区域转录的未剪接的非编码RNA序列。我们的研究结果表明sisRNAs（通过它们与调节蛋白的相互作用）具有有趣的作用，并为未剪接的稳定非编码RNA的存在提供了进一步的证据。
