a Department of Cell Biology and Molecular Genetics , University of Maryland - College Park , College Park , MD , USA.
b Basic Science Program, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research , Frederick , MD , USA.
RNA Biol. 2017 Nov 2;14(11):1466-1472. doi: 10.1080/15476286.2017.1325069. Epub 2017 Oct 11.
How plus-strand [+]RNA virus genomes transition from translation templates to replication templates is a matter of much speculation. We have previously proposed that, for Turnip crinkle virus, binding of the encoded RNA-dependent RNA polymerase (RdRp) to the 3'UTR of the [+]RNA template promotes a regional wide-spread conformational switch to an alternative structure that disassembles the cap-independent translation enhancer (CITE) in the 3'UTR. The active 3'CITE folds into a tRNA-like T-shaped structure (TSS) that binds to 80S ribosomes and 60S subunits in the P-site. In this Point-of-View, we discuss the history of our research on the TSS and our recent report combining coarse level single molecule force spectroscopy (optical tweezers) with fine-grain computer simulations of this experimental process and biochemical approaches to obtain a detailed understanding of how RdRp binding in the TSS vicinity might lead to an extensive rearrangement of the RNA structure.
正链[+]RNA 病毒基因组如何从翻译模板转变为复制模板一直是人们猜测的问题。我们之前曾提出，对于芜菁花叶病毒，编码的 RNA 依赖性 RNA 聚合酶（RdRp）与[+]RNA 模板的 3'UTR 结合，可促进局部广泛的构象转变，形成另一种结构，从而使 3'UTR 中的无帽依赖翻译增强子（CITE）解体。活性 3'CITE 折叠成类似于 tRNA 的 T 形结构（TSS），与 80S 核糖体和 P 位中的 60S 亚基结合。在本观点中，我们讨论了我们对 TSS 研究的历史，以及我们最近的报告，该报告将粗粒度单分子力光谱学（光学镊子）与该实验过程的细粒度计算机模拟以及生化方法相结合，以详细了解 RdRp 在 TSS 附近的结合如何导致 RNA 结构的广泛重排。
