GTPase activation of elongation factor EF-Tu by the ribosome during decoding

被引：154

作者：

Schuette, Jan-Christian ^{[2
]}

Murphy, Frank V. ^{[1
]}

Kelley, Ann C. ^{[1
]}

Weir, John R. ^{[1
]}

Giesebrecht, Jan ^{[2
]}

Connell, Sean R. ^{[2
]}

Loerke, Justus ^{[3
]}

Mielke, Thorsten ^{[3
]}

Zhang, Wei ^{[4
]}

Penczek, Pawel A. ^{[4
]}

Ramakrishnan, V. ^{[1
]}

Spahn, Christian M. T. ^{[2
]}

机构：

[1] MRC Lab Mol Biol, Struct Studies Div, Cambridge CB2 0QH, England

[2] Charite Univ Med Berlin, Inst Med Phys & Biophys, Berlin, Germany

[3] Max Planck Inst Mol Genet, UltraStrukturNetzwerk, Berlin, Germany

[4] Univ Texas Houston, Sch Med, Dept Biochem & Mol Biol, Houston, TX USA

来源：

EMBO JOURNAL | 2009年 / 28卷 / 06期

基金：

英国医学研究理事会; 英国惠康基金;

关键词：

cryo-electron microscopy; elongation factor; GTPase; ribosome; translation; AMINOACYL-TRANSFER-RNA; CRYSTAL-STRUCTURE; CONFORMATIONAL-CHANGE; ANGSTROM RESOLUTION; ELECTRON-MICROSCOPY; TERNARY COMPLEX; MESSENGER-RNA; ACTIVE-ROLE; SELECTION; HYDROLYSIS;

D O I：

10.1038/emboj.2009.26

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

We have used single-particle reconstruction in cryo-electron microscopy to determine a structure of the Thermus thermophilus ribosome in which the ternary complex of elongation factor Tu (EF-Tu), tRNA and guanine nucleotide has been trapped on the ribosome using the antibiotic kirromycin. This represents the state in the decoding process just after codon recognition by tRNA and the resulting GTP hydrolysis by EF-Tu, but before the release of EF-Tu from the ribosome. Progress in sample purification and image processing made it possible to reach a resolution of 6.4A. Secondary structure elements in tRNA, EF-Tu and the ribosome, and even GDP and kirromycin, could all be visualized directly. The structure reveals a complex conformational rearrangement of the tRNA in the A/T state and the interactions with the functionally important switch regions of EF-Tu crucial to GTP hydrolysis. Thus, the structure provides insights into the molecular mechanism of signalling codon recognition from the decoding centre of the 30S subunit to the GTPase centre of EF-Tu.

引用

页码：755 / 765

页数：11

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