一种16S rRNA - tRNA产物，含有一个核苷酸光三聚体，且对大肠杆菌核糖体中处于P/E杂交状态的tRNA具有特异性。
A 16S rRNA-tRNA product containing a nucleotide phototrimer and specific for tRNA in the P/E hybrid state in the Escherichia coli ribosome.
作者信息
Huggins Wayne, Wollenzien Paul
机构信息
Department of Molecular and Structural Biochemistry, North Carolina State University, Raleigh, NC 27695-7622, USA.
出版信息
Nucleic Acids Res. 2004 Dec 14;32(22):6548-56. doi: 10.1093/nar/gkh1001. Print 2004.
Ribosome complexes containing deacyl-tRNA1(Val) or biotinylvalyl-tRNA1(Val) and an mRNA analog have been irradiated with wavelengths specific for activation of the cmo5U nucleoside at position 34 in the tRNA1(Val) anticodon loop. The major product for both types of tRNA is the cross-link between 16S rRNA (C1400) and the tRNA (cmo5U34) characterized already by Ofengand and his collaborators [Prince et al. (1982) Proc. Natl Acad. Sci. USA, 79, 5450-5454]. However, in complexes containing deacyl-tRNA1(Val), an additional product is separated by denaturing PAGE and this is shown to involve C1400 and m5C967 of 16S rRNA and cmo5U34 of the tRNA. Puromycin treatment of the biotinylvalyl-tRNA1(Val) -70S complex followed by irradiation, results in the appearance of the unusual photoproduct, which indicates an immediate change in the tRNA interaction with the ribosome after peptide transfer. These results indicate an altered interaction between the tRNA anticodon and the 30S subunit for the tRNA in the P/E hybrid state compared with its interaction in the classic P/P state.
含有脱酰基 - tRNA1(Val) 或生物素化缬氨酰 - tRNA1(Val) 以及一种mRNA类似物的核糖体复合物，已用特定波长进行照射，该波长可激活tRNA1(Val) 反密码子环中第34位的cmo5U核苷。这两种类型的tRNA的主要产物是16S rRNA（C1400）与tRNA（cmo5U34）之间的交联，Ofengand及其合作者已经对其进行了表征[Prince等人（1982年），《美国国家科学院院刊》，79，5450 - 5454]。然而，在含有脱酰基 - tRNA1(Val) 的复合物中，通过变性聚丙烯酰胺凝胶电泳分离出一种额外的产物，结果表明它涉及16S rRNA的C1400和m5C967以及tRNA的cmo5U34。用嘌呤霉素处理生物素化缬氨酰 - tRNA1(Val) - 70S复合物后再进行照射，会出现这种不寻常的光产物，这表明在肽转移后tRNA与核糖体的相互作用立即发生了变化。这些结果表明，处于P/E杂交状态的tRNA与其处于经典P/P状态时相比，其反密码子与30S亚基之间的相互作用发生了改变。
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