Supplemental Data for Figs 1, 3, 4 and 6 for PLOS One Paper: HIV-1 and human PEG10 frameshift elements are functionally distinct and distinguished by novel small molecule modulators

<strong>These files are the raw data files for the PLOS One paper:</strong> <strong>HIV-1 and human PEG10 frameshift elements are functionally distinct and distinguished by novel small molecule modulators</strong> <strong>Abstract</strong> Frameshifting during translation of viral or in rare cases cellular mRNA results in the synthesis of proteins from two overlapping reading frames within the same mRNA. In HIV-1 the protease, reverse transcriptase, and integrase enzymes are in a second reading frame relative to the structural group-specific antigen (gag), and their synthesis is dependent upon frameshifting. This ensures that a strictly regulated ratio of structural proteins and enzymes, which is critical for HIV-1 replication and viral infectivity, is maintained during protein synthesis. The frameshift element in HIV-1 RNA is an attractive target for the development of a new class of anti HIV-1 drugs. However, a number of examples are now emerging of human genes using −1 frameshifting, such as PEG10 and CCR5. In this study we have compared the HIV-1 and PEG10 frameshift elements and shown they have distinct functional characteristics. Frameshifting occurs at several points within each element. Moreover, frameshift modulators that were isolated by high-throughput screening of a library of 114,000 lead-like compounds behaved differently with the PEG10 frameshift element. The most effective compounds affecting the HIV-1 element enhanced frameshifting by 2.5-fold at 10 mM in two different frameshift reporter assay systems. HIV-1 protease:gag protein ratio was affected by a similar amount in a specific assay of virally-infected cultured cell, but the modulation of frameshifting of the first-iteration compounds was not sufficient to show significant effects on viral infectivity. Importantly, two compounds did not affect frameshifting with the human PEG10 element, while one modestly inhibited rather than enhanced frameshifting at the human element. These studies indicate that frameshift elements have unique characteristics that may allow targeting of HIV-1 and of other viruses specifically for development of antiviral therapeutic molecules without effect on human genes like PEG10 that use the same generic mechanism.

本数据集为发表于《PLOS ONE》的论文《HIV-1与人类PEG10移码元件功能各异且可通过新型小分子调节剂加以区分》的原始数据文件。
【摘要】在病毒（极少数情况下为细胞）mRNA的翻译过程中发生的移码现象，可使同一条mRNA内的两个重叠阅读框均可合成蛋白质。在HIV-1中，蛋白酶、逆转录酶与整合酶的编码序列位于结构特异性抗原（group-specific antigen, gag）阅读框的下游第二阅读框内，其合成依赖于移码过程。这确保了在蛋白质合成过程中，结构蛋白与酶类的比例受到严格调控——该比例对于HIV-1的复制与病毒感染性至关重要。HIV-1 RNA中的移码元件，是开发新型抗HIV-1药物的极具潜力的靶点。但目前已有多项研究表明，人类基因中也存在利用-1移码机制的案例，例如PEG10与CCR5。本研究对HIV-1与PEG10的移码元件进行了对比分析，证实二者具备截然不同的功能特性。两类元件内部均可在多个位点发生移码现象。此外，通过对114000个类先导化合物库进行高通量筛选得到的移码调节剂，对PEG10移码元件的作用效果与对HIV-1移码元件存在显著差异。在两种不同的移码报告基因检测系统中，针对HIV-1移码元件效果最优的化合物可在10mM浓度下将移码效率提升2.5倍。在病毒感染的培养细胞特异性检测实验中，HIV-1蛋白酶与gag蛋白的比例也受到了相似幅度的调控，但初代化合物对移码过程的调控强度不足以对病毒感染性产生显著影响。值得注意的是，有两种化合物对人类PEG10移码元件的移码过程无任何影响，而另有一种化合物仅会轻度抑制而非提升该元件的移码效率。本研究结果表明，各类移码元件具备独特的序列与功能特性，这使得我们可以针对性地靶向HIV-1及其他病毒，开发抗病毒治疗分子，而不会对PEG10这类利用相同通用移码机制的人类基因产生影响。