Labeling the peptidyltransferase center of the Escherichia coli ribosome with photoreactive tRNA(Phe) derivatives containing azidoadenosine at the 3' end of the acceptor arm: a model of the tRNA-ribosome complex.
Labeling the peptidyltransferase center of the Escherichia coli ribosome with photoreactive tRNA(Phe) derivatives containing azidoadenosine at the 3' end of the acceptor arm: a model of the tRNA-ribosome complex.
复制标题
用受体臂 3 端含有叠氮腺苷的光反应 tRNA(Phe) 衍生物标记大肠杆菌核糖体的肽基转移酶中心：tRNA-核糖体复合物的模型。
DOI：
10.1073/pnas.86.14.5232
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发表时间：
1989
影响因子：
11.1
通讯作者：
Zimmermann,RA
中科院分区：
文献类型：
--
作者：
Wower,J;Hixson,SS;Zimmermann,RA
在73或76位含有2-叠氮腺苷（2N 3A）的酵母tRNA（Phe）的光反应衍生物已交联到大肠杆菌核糖体的肽基位点。共价tRNA-核糖体附着依赖于tRNA中腺苷被2N 3A取代、300 nm光照射和poly（U）的存在。在所有情况下，修饰的tRNA都只与50 S核糖体亚基交联。在第73位含有2N 3A的tRNA衍生物仅标记蛋白质L27，而在第76位含有2N 3A的tRNA衍生物标记蛋白质L15、L16和L27以及23 S rRNA的片段。rRNA中的交联位点被鉴定为鸟苷-1945，其位于与许多修饰的碱基相邻的高度保守序列内，并且直到现在才在肽基转移酶中心被鉴定。基于这些结果，以及之前报道的3 '末端-A-C-C-A序列中含有8-叠氮腺苷的tRNA的交联[Wower，J.，希克森，S. S. & Zimmermann，R. A.（1988）Biochemistry 27，8114-8121]，我们提出了tRNA分子在肽基和氨酰基位点的排列模型，该模型与关于肽基转移酶中心和E.大肠杆菌核糖体。
Photoreactive derivatives of yeast tRNA(Phe) containing 2-azidoadenosine (2N3A) at position 73 or 76 have been crosslinked to the peptidyl site of Escherichia coli ribosomes. Covalent tRNA-ribosome attachment was dependent upon the replacement of adenosine by 2N3A in the tRNA, irradiation with 300-nm light, and the presence of poly(U). In all cases, the modified tRNAs became crosslinked exclusively to 50S ribosomal subunits. While the tRNA derivative containing 2N3A at position 73 labeled only protein L27, that containing 2N3A at position 76 labeled proteins L15, L16, and L27 as well as a segment of the 23S rRNA. The site of crosslinking in the rRNA was identified as guanosine-1945, which lies within a highly conserved sequence adjacent to a number of modified bases and has not until now been identified at the peptidyltransferase center. On the basis of these results, and previously reported crosslinks from tRNA containing 8-azidoadenosine in the 3'-terminal -A-C-C-A sequence [Wower, J., Hixson, S. S. & Zimmermann, R. A. (1988) Biochemistry 27, 8114-8121], we propose a model for the arrangement of tRNA molecules at the peptidyl and aminoacyl sites that is consistent with most of the information available about the location of the peptidyltransferase center and the decoding domain of the E. coli ribosome.