Martin R P, Sibler A P, Schneller J M, Keith G, Stahl A J, Dirheimer G
Nucleic Acids Res. 1978 Dec;5(12):4579-92. doi: 10.1093/nar/5.12.4579.
Mitochondrial tRNAPhe from Saccharomyces cerevisiae isolated by two-dimensional gel electrophoresis was sequenced by fingerprinting uniformly labeled 32 P-tRNA as well as by 5'-end postlabeling techniques. Its sequence was found to be: pG-C-U-U-U-U-A-U-A-G-C-U-U-A-G-D-G-G-D-A-A-A-G-C-m22G-A-U-A-A-A-phi-U-G-A-A-m1G-A-phi-U-U-A-U-U-U-A-C-A-U-G-U-A-G-U-phi-C-G-A-U-U-C-U-C-A-U-U-A-A-G-G-G-C-A-C-C-A. The secondary structure we propose, in order to maximize base pairing in the phiC stem and to allow tertiary interaction between G15 and C46, excludes U50 from base pairing giving a bulge in the phiC stem. No conclusion can be drawn concerning the endosymbiotic theory of mitochondria evolution by comparing the primary structure of mt. tRNAPhe with other sequenced tRNAsPhe. This mt.tRNAPhe lacks some of the structural elements reported to be involved in the yeast cytoplasmic phenylalanyl-tRNA ligase recognition site and cannot be aminoacylated by purified yeast cytoplasmic phenylalanyl-tRNA ligase.
通过二维凝胶电泳从酿酒酵母中分离出的线粒体苯丙氨酸转运RNA(tRNAPhe),采用对均匀标记的32P - tRNA进行指纹图谱分析以及5'端后标记技术进行测序。其序列如下:pG - C - U - U - U - U - A - U - A - G - C - U - U - A - G - D - G - G - D - A - A - A - G - C - m22G - A - U - A - A - A - phi - U - G - A - A - m1G - A - phi - U - U - A - U - U - U - A - C - A - U - G - U - A - G - U - phi - C - G - A - U - U - C - U - C - A - U - U - A - A - G - G - G - C - A - C - C - A。为了使phiC茎中的碱基对最大化,并允许G15和C46之间的三级相互作用,我们提出的二级结构将U50排除在碱基对之外,从而在phiC茎中形成一个凸起。通过比较线粒体tRNAPhe与其他已测序的tRNAPhe的一级结构,无法得出关于线粒体进化内共生理论的结论。这种线粒体tRNAPhe缺乏一些据报道参与酵母细胞质苯丙氨酰 - tRNA连接酶识别位点的结构元件,并且不能被纯化的酵母细胞质苯丙氨酰 - tRNA连接酶氨酰化。