Fouts D E, Celander D W
Department of Microbiology, University of Illinois at Urbana-Champaign, Urbana, IL, USA.
Gene. 1998 Mar 27;210(1):135-42. doi: 10.1016/s0378-1119(98)00066-3.
The RNA challenge phage system enables genetic selection of proteins with RNA-binding activity in bacteria. These phages are modified versions of the temperate DNA bacteriophage P22 in which post-transcriptional regulatory events control the developmental fate of the phage. The system was originally developed to identify novel RNA ligands that display reduced affinity for the R17/MS2 coat protein, as well as to select for suppressor coat proteins that recognize mutant RNA ligands. During the course of evaluating whether the HIV-1 Rev protein could direct lysogen development for bacteriophage derivatives that encode Rev response element (RRE) RNA sequences, two examples of RRE RNA ligands that interfere with challenge phage development were identified. In the phage examples described, RRE RNA secondary structure prevents Ant protein biosynthesis and lytic development. Phage lysogen formation occurs efficiently in recipient cells, independent of the expression status of the Rev protein or trans-acting competitor RRE RNA ligands. These studies provide the first example whereby RNA challenge phages may be applied to study RNA folding events and RNA structural interactions in an in vivo context.
RNA 挑战噬菌体系统能够在细菌中对具有 RNA 结合活性的蛋白质进行遗传筛选。这些噬菌体是温和型 DNA 噬菌体 P22 的修饰版本，其中转录后调控事件控制着噬菌体的发育命运。该系统最初是为了鉴定对 R17/MS2 外壳蛋白亲和力降低的新型 RNA 配体，以及筛选识别突变 RNA 配体的抑制性外壳蛋白而开发的。在评估 HIV-1 Rev 蛋白是否能够指导编码 Rev 反应元件（RRE）RNA 序列的噬菌体衍生物的溶原发育过程中，发现了两个干扰挑战噬菌体发育的 RRE RNA 配体实例。在所描述的噬菌体实例中，RRE RNA 的二级结构阻止了 Ant 蛋白的生物合成和裂解发育。噬菌体溶原形成在受体细胞中高效发生，与 Rev 蛋白或反式作用竞争性 RRE RNA 配体的表达状态无关。这些研究首次证明了 RNA 挑战噬菌体可用于在体内研究 RNA 折叠事件和 RNA 结构相互作用。
