EttA regulates translation by binding the ribosomal E site and restricting ribosome-tRNA dynamics.

EttA regulates translation by binding the ribosomal E site and restricting ribosome-tRNA dynamics.
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DOI:
10.1038/nsmb.2741
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发表时间:
2014-02
影响因子:
16.8
通讯作者:
Frank J
Frank J
中科院分区:
生物学1区
文献类型:
--
作者:
Chen B;Boël G;Hashem Y;Ning W;Fei J;Wang C;Gonzalez RL Jr;Hunt JF;Frank J

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细胞表达许多核糖体相互作用因子,其功能和分子机制仍然未知。在这里,我们阐明了新表征的调节翻译因子能量依赖性翻译节流阀 A (EttA) 的机制,它是 ATP 结合盒 F (ABC-F) 蛋白家族的大肠杆菌代表。使用冷冻电镜,我们证明 EttA 的 ATP 结合形式与核糖体 tRNA 出口 (E) 位点结合,在核糖体 L1 茎和肽基-tRNA 结合 (P) 位点中结合的 tRNA 之间形成桥接相互作用。使用单分子荧光共振能量转移 (smFRET),我们发现 EttA 的 ATP 结合形式限制了蛋白质合成所需的核糖体和 tRNA 动力学。据我们所知,这项工作代表了第一个例子,其中任何 ABC-F 家族蛋白的详细分子机制已被确定,并建立了一个框架来阐明其他调节翻译因子的机制。
Cells express many ribosome-interacting factors whose functions and molecular mechanisms remain unknown. Here, we elucidate the mechanism of a newly characterized regulatory translation factor, Energy-dependent Translational Throttle A (EttA), which is an Escherichia coli representative of the ATP-binding cassette F (ABC-F) protein family. Using cryo-EM, we demonstrate that the ATP-bound form of EttA binds to the ribosomal tRNA exit (E) site, where it forms bridging interactions between the ribosomal L1 stalk and the tRNA bound in the peptidyl-tRNA binding (P) site. Using single-molecule fluorescence resonance energy transfer (smFRET), we show that the ATP-bound form of EttA restricts ribosome and tRNA dynamics required for protein synthesis. This work represents the first example, to our knowledge, where the detailed molecular mechanism of any ABC-F family protein has been determined and establishes a framework for elucidating the mechanisms of other regulatory translation factors.
DOI: 10.1038/nsmb.2740
发表时间: 2014-02
影响因子: 16.8
作者:
Boël G;Smith PC;Ning W;Englander MT;Chen B;Hashem Y;Testa AJ;Fischer JJ;Wieden HJ;Frank J;Gonzalez RL Jr;Hunt JF
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