Nature2014-06-27 1:18 PM

“摇晃”的碱基对决定tRNA的保真度 The selective tRNA aminoacylation mechanism based on a single G•U pair

论文摘要 

tRNA合成酶必须将合适的氨基酸附着到合适的、同源的tRNA上来维持遗传代码向蛋白质的正确翻译。丙氨酸的tRNA中一个“非华生-克里克碱基对”G3•U70确保被合成酶(AlaRS)正确氨酰化。Shigeyuki Yokoyama及同事报告了来自古菌Archaeoglobus fulgidus、在与携带G3•U70的tRNAAla(tRNAAla/GU)和与携带 A3•U70的一个变体(tRNAAla/AU)以及与一个alanyl-AMP类似物形成的复合物中的AlaRS的两个晶体结构。这两个结构的比较显示了该合成酶具有选择性的基础。G3•U70的独特几何决定 tRNA的CCA区域的取向,引导末端腺苷进入催化点。这一机制可解释远离该催化点的少量核苷酸何以能决定特定氨酰化的反应性。

Abstract 

Ligation of tRNAs with their cognate amino acids, by aminoacyl-tRNA synthetases, establishes the genetic code. Throughout evolution, tRNAAla selection by alanyl-tRNA synthetase (AlaRS) has depended predominantly on a single wobble base pair in the acceptor stem, G3•U70, mainly on the kcat level. Here we report the crystal structures of an archaeal AlaRS in complex with tRNAAla with G3•U70 and its A3•U70 variant. AlaRS interacts with both the minor- and the major-groove sides of G3•U70, widening the major groove. The geometry difference between G3•U70 and A3•U70 is transmitted along the acceptor stem to the 3′-CCA region. Thus, the 3′-CCA region of tRNAAla with G3•U70 is oriented to the reactive route that reaches the active site, whereas that of the A3•U70 variant is folded back into the non-reactive route. This novel mechanism enables the single wobble pair to dominantly determine the specificity of tRNA selection, by an approximate 100-fold difference in kcat.

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