Giegé R, Briand J P, Mengual R, Ebel J P, Hirth L
Eur J Biochem. 1978 Mar;84(1):251-6. doi: 10.1111/j.1432-1033.1978.tb12163.x.
A comparative study of the aminoacylation of the two RNA components of turnip yellow mosaic virus, of yeast tRNAVal, tRNAfMet and of tRNAPhe by purified yeast valyl-tRNA synthetase is reported. Aminoacylations were performed in the presence of pure yeast tRNA nucleotidyltransferase, since 85% of the viral RNA molecules lacked the 3'-adenosine. We find that aminoacylation of the viral RNAs, like tRNA aminoacylation, reflects an equilibrium between the acylation and deacylation reactions. The kinetic parameters of TYM virus RNA valylation resemble the values found for tRNAVal valylation; in particular, there is a strong affinity between the viral RNA and valyl-tRNA synthetase and the rate constant for TYM virus RNA valylation is only slightly lower than that for tRNAVal. This result contrasts with the reduced rates observed in tRNA mischarging, and suggests that the viral RNA could be easily aminoacylated in vivo. Considering the fact that the 3'-terminal sequence of TYM virus RNA has only a few points of resemblance to a tRNA sequence, we propose that there are some structural motifs found in both tRNAVal and TYM virus RNA which are brought in a similar spatial arrangement recognized by valyl-tRNA synthetase.
本文报道了用纯化的酵母缬氨酰 - tRNA合成酶对芜菁黄花叶病毒的两种RNA组分、酵母tRNAVal、tRNAfMet和tRNAPhe进行氨酰化的比较研究。由于85%的病毒RNA分子缺少3'-腺苷,氨酰化反应在纯酵母tRNA核苷酸转移酶存在的情况下进行。我们发现,病毒RNA的氨酰化,与tRNA氨酰化一样,反映了酰化反应和去酰化反应之间的平衡。芜菁黄花叶病毒RNA缬氨酰化的动力学参数类似于tRNAVal缬氨酰化的参数;特别是,病毒RNA与缬氨酰 - tRNA合成酶之间有很强的亲和力,芜菁黄花叶病毒RNA缬氨酰化的速率常数仅略低于tRNAVal的速率常数。这一结果与tRNA错配时观察到的速率降低形成对比,并表明病毒RNA在体内可能很容易被氨酰化。考虑到芜菁黄花叶病毒RNA的3'-末端序列与tRNA序列只有几个相似点,我们提出在tRNAVal和芜菁黄花叶病毒RNA中都存在一些结构基序,它们以相似的空间排列方式被缬氨酰 - tRNA合成酶识别。