X-ray crystallography of maturation and aminoacylation of transfer RNA

转移 RNA 成熟和氨酰化的 X 射线晶体学

• 批准号: 14580668
• 负责人: NUREKI Osamu
• 金额: $ 2.3万
• 依托单位: Tokyo Institute of Technology (2003)The University of Tokyo (2002)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14580668/
• 关键词: X-ray crystallography; transfer RNA; RNA processing; post-transcriptional modification; aminoacyl-tRNA synthetase; apoptosis; 遺伝情報変換; タンパク質合成; トランスファーRNA; RNAの転写後修飾

We solved crystal structures of several enzymes involved in tRNA maturation and post-transcriptional modification and their complex with tRNA. In the processing step, 3'-extended trailer sequence is trimmed by RNase PH, and, if RNase PH trims off a part of the amino-acid-attaching CCA terminus of tRNA, then CCA-adding enzyme (template-independent RNA polymerase) repairs the CCA sequence. Crystal structure of Aquifex aeolicus RNase PH at 2.3 Å resolution) reveals how two enzyme families have evolved to polymerize defined CCA trinucleotides without aid of any nucleic-acid template. In the post-transcriptional modification process, crystal structure (3.3 Å resolution) of archaeosine tRNA guanine transglycosylase in a complex with tRNA uncovered how the enzyme recognizes and modifies a deeply-buried nucleotide by deformation of canonical tRNA L-shape to 1 -shape. The 1.85 Å-resolution crystal structure of tRNA(gm18)methyltransferase revealed that the deep knot structure found in the catalytic fold not only constructs active-site and cofactor-binding site but also facilitates RNA-dependent methylation mechanism via molecular dimerization. For aminoacyl-tRNA synthetase, we elucidated the structural basis for the cytokine activity of human tryptophanyl-tRNA synthetase by crystallographic analysis, which has been gained in the long evolution of the enzyme.

我们解决了几个参与tRNA成熟和转录后修饰的酶的晶体结构，以及它们与tRNA的复合物。在加工步骤中，通过RNase PH修剪3 ′延伸的尾部序列，并且如果RNase PH修剪掉tRNA的氨基酸连接CCA末端的一部分，则CCA添加酶（模板非依赖性RNA聚合酶）修复CCA序列。Aquifex aeolicus RNase PH的晶体结构（2.3 μ m分辨率）揭示了两种酶家族如何在没有任何核酸模板的帮助下进化为单克隆确定的CCA三核苷酸。在转录后修饰过程中，与tRNA复合的古核苷tRNA鸟嘌呤转糖基酶的晶体结构（3.3 μ m分辨率）揭示了该酶如何通过将典型的tRNA L形变形为1形来识别和修饰深埋的核苷酸。tRNA（gm 18）甲基转移酶1.85 nm分辨率的晶体结构显示，在催化折叠中发现的深结结构不仅构建了活性位点和辅因子结合位点，而且通过分子二聚化促进了RNA依赖的甲基化机制。对于氨酰-tRNA合成酶，我们通过晶体学分析阐明了人氨酰-tRNA合成酶细胞因子活性的结构基础，这是在酶的长期进化中获得的。

项目成果

R.Ishii, O.Nureki, S.Yokoyama: "Crystal structure of the tRNA processing enzyme RNase PH from Aquifex aeolicus."J.Biol. Chem.. 278. 32397-32404 (2003)

R.Ishii、O.Nureki、S.Yokoyama：“来自 Aquifex aeolicus 的 tRNA 加工酶 RNase PH 的晶体结构。”J.Biol。

S.Sekine, O.Nureki et al.: "Two alternative ATP-binding modes mediate tRNA-dependent activation of Glu-tRNA synthetase"EMBO J.. 23. 1-13 (2002)

S.Sekine、O.Nureki 等人：“两种替代的 ATP 结合模式介导 Glu-tRNA 合成酶的 tRNA 依赖性激活”EMBO J.. 23. 1-13 (2002)

M.Okabe, K.Tomita, O.Nureki^*, S.Yokoyama et al.(^*correspondence): "Divergent evolution of trinucleotide polymerization revealed by an archaeal CCA-adding enzyme structure."EMBO J.. 22. 5918-5927 (2003)

M.Okabe、K.Tomita、O.Nureki^*、S.Yokoyama 等人（^*通讯）：“古细菌 CCA 添加酶结构揭示的三核苷酸聚合的发散进化。”EMBO J.. 22. 5918

Y.Kise, S.W.Lee, S.G.Park, S.Fukai, S.Kim, O.Nureki: "A short peptide insertion crucial for angiostatic activity of human tryptophanyl-tRNA synthetase."Nat.Struct. & Mol.Biol.. 11. 149-156 (2004)

Y.Kise、S.W.Lee、S.G.Park、S.Fukai、S.Kim、O.Nureki：“短肽插入对于人色氨酰-tRNA 合成酶的血管抑制活性至关重要。”Nat.Struct。

Y.Kise, S.W.Lee, S.G.Park, S.Fukai, S.Kim, O.Nureki: "A short peptide insertion crucial for angiostatic activity of human tryptophanyl-tRNA synthetase."Nat.Struct.& Mol.Biol.. 11. 149-156 (2004)

Y.Kise、S.W.Lee、S.G.Park、S.Fukai、S.Kim、O.Nureki：“短肽插入对于人色氨酰-tRNA 合成酶的血管抑制活性至关重要。”Nat.Struct。