Briand J P, Jonard G, Guilley H, Richards K, Hirth L
Eur J Biochem. 1977 Feb;72(3):453-63. doi: 10.1111/j.1432-1033.1977.tb11269.x.
The experiments described in this paper and the following one establish the sequence of the 3'-OH terminal 159 nucleotides of turnip yellow mosaic virus RNA. Uniformly 32P-labeled turnip yellow mosaic virus RNA was partially digested with T1 ribonuclease and the fragments were fractionated by polyacrylamide gel electrophoresis. Fragments originating from the 3'-OH end of the RNA molecule were identified by testing for the 3'-terminal oligonucleotide, C-COH, after total U2 ribonuclease hydrolysis. Once identified, the 3'-OH terminal fragments were sequenced by the methods of Sanger et al. The first 51 nucleotides of the longest of the sequenced fragments (158 nucleotides) extends into the 3'-terminal part of the coat protein cistron. The coat protein cistron is followed by a stretch of 108 untranslated nucleotides whose function, though still unknown, is probably linked to the tRNA-like properties which have been attributed to the 3'-OH extremity of this viral RNA. Two possible secondary structures are proposed for the sequence and the implications of the findings with regard to the tRNA-like properties of the extremity are discussed.
本文及后续文章中所描述的实验确定了芜菁黄花叶病毒RNA 3'-OH末端159个核苷酸的序列。用T1核糖核酸酶对均匀标记32P的芜菁黄花叶病毒RNA进行部分消化,然后通过聚丙烯酰胺凝胶电泳对片段进行分级分离。通过在完全用U2核糖核酸酶水解后检测3'-末端寡核苷酸C-COH,鉴定出源自RNA分子3'-OH末端的片段。一旦鉴定出来,就通过桑格等人的方法对3'-OH末端片段进行测序。所测序片段中最长的片段(158个核苷酸)的前51个核苷酸延伸到外壳蛋白顺反子的3'-末端部分。外壳蛋白顺反子之后是一段108个未翻译的核苷酸,其功能虽然仍然未知,但可能与该病毒RNA 3'-OH末端所具有的类似tRNA的特性有关。针对该序列提出了两种可能的二级结构,并讨论了这些发现对于末端类似tRNA特性的意义。