Two alternative conformations of mRNA in the human ribosome during elongation and termination of translation as revealed by EPR spectroscopy,Computational and Structural Biotechnology Journal

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Two alternative conformations of mRNA in the human ribosome during elongation and termination of translation as revealed by EPR spectroscopy
Computational and Structural Biotechnology Journal ( IF 4.4 ) Pub Date : 2021-08-19 , DOI: 10.1016/j.csbj.2021.08.024
Konstantin N Bulygin ₁ , Ivan O Timofeev ₂ , Alexey A Malygin ₁ , Dmitri M Graifer ₁ , Maria I Meschaninova ₁ , Alya G Venyaminova ₁ , Olesya A Krumkacheva ₂ , Matvey V Fedin ₂ , Ludmila Yu Frolova ₃ , Galina G Karpova ₁ , Elena G Bagryanskaya ₄

Affiliation

The conformation of mRNA in the region of the human 80S ribosome decoding site was monitored using 11-mer mRNA analogues that bore nitroxide spin labels attached to the terminal nucleotide bases. Intramolecular spin–spin distances were measured by DEER/PELDOR spectroscopy in model complexes mimicking different states of the 80S ribosome during elongation and termination of translation. The measurements revealed that in all studied complexes, mRNA exists in two alternative conformations, whose ratios are different in post-translocation, pre-translocation and termination complexes. We found that the presence of a tRNA molecule at the ribosomal A site decreases the relative share of the more extended mRNA conformation, whereas the binding of eRF1 (alone or in a complex with eRF3) results in the opposite effect. In the termination complexes, the ratios of mRNA conformations are practically the same, indicating that a part of mRNA bound in the ribosome channel does not undergo significant structural alterations in the course of completion of the translation. Our results contribute to the understanding of mRNA molecular dynamics in the mammalian ribosome channel during translation.

中文翻译：

EPR 光谱揭示了人类核糖体中 mRNA 在翻译延伸和终止过程中的两种替代构象

使用 11 聚体 mRNA 类似物监测人 80S 核糖体解码位点区域中 mRNA 的构象，该类似物带有附着在末端核苷酸碱基上的硝基氧自旋标签。通过模拟 80S 核糖体在延伸和翻译终止过程中的不同状态的模型复合物中的 DEER/PELDOR 光谱来测量分子内自旋-自旋距离。测量结果表明，在所有研究的复合物中，mRNA以两种替代构象存在，其比例在易位后、易位前和终止复合物中不同。我们发现核糖体 A 位点上 tRNA 分子的存在降低了更延伸的 mRNA 构象的相对份额，而 eRF1 的结合（单独或与 eRF3 形成复合物）会产生相反的效果。在终止复合物中，mRNA构象的比例实际上是相同的，这表明结合在核糖体通道中的部分mRNA在翻译完成的过程中没有发生显着的结构改变。我们的结果有助于理解翻译过程中哺乳动物核糖体通道中 mRNA 分子动力学。

更新日期：2021-08-19

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