Institute of Protein Research, Russian Academy of Sciences, Institutskaya 4, Pushchino, Moscow Region 142290, Russian Federation.
Institute of Protein Research, Russian Academy of Sciences, Institutskaya 4, Pushchino, Moscow Region 142290, Russian Federation.
J Struct Biol. 2020 Sep 1;211(3):107559. doi: 10.1016/j.jsb.2020.107559. Epub 2020 Jul 10.
Complexes of archaeal ribosomal proteins uL11 and uL10/P0 (the two-domain N-terminal fragment of uL10, uL10NTF/P0NTF) with the adjacent 74 nucleotides of 23S rRNA fragment (23SrRNA(74)) from Methanococcus jannaschii (Mja) were obtained, crystallized and their structures were studied. The comparative structural analysis of the complexes of Mja uL10NTF•23SrRNA(74) and Mja uL10NTF•uL11•23SrRNA(74) shows that the insertion of uL11 in the binary complex does not change the conformation of the 23S rRNA fragment. On the other hand, the interaction with this specific RNA fragment leads to the restructuring of uL11 compared to the structure of this protein in the free state. Besides, although analysis confirmed the mobility of uL10/P0 domain II, disproved the assumption that it may be in contact with rRNA or uL11. In addition, the Mja uL10NTF•uL11•23SrRNA(74) complex was cocrystallized with the antibiotic thiostrepton, and the structure of this complex was solved. The thiostrepton binding site in this archaeal complex was found between the 23S rRNA and the N-terminal domain (NTD) of the Mja uL11 protein, similar to its binding site in the one of bacterial ribosome complex with thiostrepton. Upon binding of thiostrepton, the NTD of uL11 shifts toward rRNA by 7 Å. Such a shift may be the cause of the inhibitory effect of the antibiotic on the recruitment of translation factors to the GTPase-activating region in archaeal ribosomes, similar to its inhibitory effect on protein synthesis in bacterial ribosomes.
获得了古菌核糖体蛋白 uL11 和 uL10/P0（uL10 的两个结构域 N 端片段，uL10NTF/P0NTF）与来自 Methanococcus jannaschii（Mja）的 23S rRNA 片段（23SrRNA(74)）的相邻 74 个核苷酸的复合物，并对其进行了结晶和结构研究。Mja uL10NTF·23SrRNA(74)和 Mja uL10NTF·uL11·23SrRNA(74)复合物的比较结构分析表明，uL11 的插入不会改变 23S rRNA 片段的构象。另一方面，与该特定 RNA 片段的相互作用导致 uL11 的构象与游离状态下该蛋白的结构相比发生了重构。此外，尽管分析证实了 uL10/P0 结构域 II 的可移动性，但否定了其可能与 rRNA 或 uL11 接触的假设。此外，Mja uL10NTF·uL11·23SrRNA(74) 复合物与抗生素硫链丝菌素共结晶，并解析了该复合物的结构。在这个古菌复合物中，硫链丝菌素的结合位点位于 23S rRNA 和 Mja uL11 蛋白的 N 端结构域（NTD）之间，与它在与硫链丝菌素结合的细菌核糖体复合物中的结合位点相似。硫链丝菌素结合后，uL11 的 NTD 向 rRNA 移动 7Å。这种移动可能是抗生素对古菌核糖体中 GTPase 激活区域翻译因子招募的抑制作用的原因，类似于它对细菌核糖体中蛋白质合成的抑制作用。
