The selective tRNA aminoacylation mechanism based on a single G•U pair.

The selective tRNA aminoacylation mechanism based on a single G•U pair.
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DOI:
10.1038/nature13440
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发表时间:
2014-06-26
期刊:
影响因子:
64.8
通讯作者:
Yokoyama, Shigeyuki
Yokoyama, Shigeyuki
中科院分区:
综合性期刊1区
文献类型:
--
作者:
Naganuma, Masahiro;Sekine, Shun-ichi;Chong, Yeeting Esther;Guo, Min;Yang, Xiang-Lei;Gamper, Howard;Hou, Ya-Ming;Schimmel, Paul;Yokoyama, Shigeyuki

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通过氨基酰基trna合成酶将trna与其同源氨基酸连接,建立遗传密码。在整个进化过程中,丙烯酰- trna合成酶(AlaRS)对tRNAAla的选择主要依赖于受体茎中的单个摆动碱基对G3•U70,主要依赖于kcat水平。本文报道了一种古细菌AlaRS与tRNAAla G3•U70及其A3•U70变体的配合物的晶体结构。AlaRS与G3•U70的小槽和大槽边相互作用,使大槽变宽。G3•U70和A3•U70之间的几何差异沿着受体杆传输到3 ' -CCA区域。因此,具有G3•U70的tRNAAla的3 ' -CCA区域面向到达活性位点的反应性途径,而A3•U70变体的3 ' -CCA区域则折叠回“非反应性途径”。这种新机制使得单个摆动对通过kcat大约100倍的差异来主导tRNA选择的特异性。
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 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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