2002 年度実績報告書

RNAタンパク質複合体構造

研究課題

研究課題/領域番号	14035205
研究機関	東京大学
研究代表者	横山茂之東京大学, 大学院・理学系研究科, 教授 (00159229)
研究分担者	河合剛太千葉工業大学, 工学部, 助教授 (70211860)
キーワード	X線結晶構造解析 / NMR / 立体構造 / RNA / RNA結合タンパク質 / リボゾーム / RNAポリメーゼ
研究概要	1.X線結晶構造解析法:(1)超好熱古細菌Pyrococcus horikoshiiのリジルtRNA合成酵素(LysRS)の結晶構造を決定し,構造の全く異なる2種類のLysRSが同じアミノ酸,tRNAを認識するメカニズムを明らかにした.超好熱古細菌Methanococcus jannashii由来のチロシルtRNA合成酵素(TyrRS)とtRNA^<Tyr>とL-チロシンの複合体の結晶構造を決定し,真核型TyrRSによるtRNAの認識機構を解明し,さらにTyrRS変異体によりアンバー終止コドンに非天然型アミノ酸を取り込ませた.また,大腸菌由来の変異体TyrRSと3-ヨードチロシンとの複合体,古細菌のアーケオシングアニントランスグリコシダーゼとtRNAの複合体,および,tRNAの3末端のトリミングを行うRNase PHの結晶構造を決定した.(2)リボソームRNAのメチル化を触媒する2'-O-MTaseの結晶構造を決定し,本酵素ファミリーの触媒部位には明瞭な結び目(knot)構造があることを発見した.(3)既知のRNA結合ドメインをもたないAUH蛋白質の結晶構造を決定した.(4)分子量40万からなる真正細菌のRNAポリメラーゼのホロ酵素(α_2ββ'ωσ)の結晶構造を2.6分解能で解明し,σサブユニットのN末端側の2つのドメインがV字型で配置されていること,および,そのC末端側のドメインがRNA鎖の出口付近に配置されていることを明らかにした.さらに,T7RNAポリメラーゼの伸長複合体の結晶構造を解明した。 2.NMR法:グループIイントロンのP7/P9.0領域に対応するRNAの立体構造を解析し,グアノシン残基の認識メカニズムを解明した.また,ヒトSRP RNAの一部であるhelix6に相当するRNA,および,tmRNAと結合する高度好熱菌SmpBの溶液構造を決定した.

研究成果
(18件)

すべてその他

すべて文献書誌 (18件)

[文献書誌] Sekine, S.: "ATP binding by glutamyl-tRNA synthetase is switched to the productive mode by tRNA binding"EMBO J.. 23. 676-688 (2003)
[文献書誌] Someya, T.: "Solution structure of a tmRNA-binding protein, SmpB, from Thermus thermophilus"FEBS Lett.. 535. 94-100 (2003)
[文献書誌] Fukai, S.: "Mechanism of molecular interactions for tRNA^<Val> recognition by valyl-tRNA synthetase"RNA. 9. 100-111 (2003)
[文献書誌] Temiakov, D.: "Crystallization and preliminary crystallographic analysis of T7 RNA polymerase elongation complex"Acta Cryst.. D59. 185-187 (2003)
[文献書誌] Tachirov, T.H.: "Structure of a T7 RNA polymerase elongation complex at 2.9 A resolution"Nature. 420. 43-50 (2002)
[文献書誌] Sakamoto, K.: "Site-specific incorporation of an unnatural amino acid into proteins in mammalian cells"Nucleic Acids Res. 30. 4692-4699 (2002)
[文献書誌] Vassylyeva, M.N.: "Purification, crystallization and initial crystallographic analysis of RNA polymerase holoenzyme from Thermus thermophilus"Acta Cryst.. D58. 1497-1500 (2002)
[文献書誌] Kiga, D.: "An engineered Escherichia coli tyrosyl-tRNA synthetase for site-specific incorporation of an unnatural amino acid into proteins in eukaryotic translation and its application in a wheat germ cell-free system"Proc. Natl. Acad. Sci. U.S.A.. 99. 9715-9720 (2002)
[文献書誌] Nureki, O.: "an enzyme with a deep trefoil knot for the active-site architecture"Acta Cryst.. D58. 1129-1137 (2002)
[文献書誌] Vassylyev, D.G.: "Crystal structure of a bacterial RNA polymerase holoenzyme at 2.6 A resolution"Nature. 417. 712-719 (2002)
[文献書誌] Kitamura, A.: "Solution structure of an RNA fragment with the P7/P9.0 region and the 3'-terminal guanosine of the tetrahymena group I intron"RNA. 8. 440-451 (2002)
[文献書誌] Terada, T.: "Functional convergence of two lysyl-tRNA synthetases with unrelated topologies"Nature Struct. Biol.. 9. 257-262 (2002)
[文献書誌] Kitabatake, M.: "Indolmycin resistance of Streptomyces coelicolor A3(2) by induced expression of one of its two tryptophanyl-tRNA synthetases"J. Biol. Chem.. 277. 23882-23887 (2002)
[文献書誌] Hendrickson, T.L.: "Mutational separation of two pathways for editing by a class I tRNA synthetase"Mol. Cell. 9. 353-362 (2002)
[文献書誌] Hirao, I.: "An unnatural base pair for incorporating amino acid analogs into proteins"Nature Biotechnol.. 20. 177-182 (2002)
[文献書誌] Takasu, A.: "Analysis of Relative Positions of Ribonucleotide Bases in a Crystal Structure of Ribosome"Nucleotides and Nucleic Acids. 21. 449-462 (2002)
[文献書誌] Takasu, A.: "Classification of RNA Structures Based on Hydrogen Bond and Base-Base Stacking Patterns : Application for NMR structures"J. Biochem.. 132. 211-215 (2002)
[文献書誌] Sakamoto, T: "Solution structure of a SRP19 binding domain in human SRP RNA"J. Biochem.. 132. 177-182 (2002)

2002 年度 実績報告書

RNAタンパク質複合体構造

研究代表者

横山 茂之 東京大学, 大学院・理学系研究科, 教授 (00159229)

研究成果

[文献書誌] Sekine, S.: "ATP binding by glutamyl-tRNA synthetase is switched to the productive mode by tRNA binding"EMBO J.. 23. 676-688 (2003)

[文献書誌] Someya, T.: "Solution structure of a tmRNA-binding protein, SmpB, from Thermus thermophilus"FEBS Lett.. 535. 94-100 (2003)

[文献書誌] Fukai, S.: "Mechanism of molecular interactions for tRNA^<Val> recognition by valyl-tRNA synthetase"RNA. 9. 100-111 (2003)

[文献書誌] Temiakov, D.: "Crystallization and preliminary crystallographic analysis of T7 RNA polymerase elongation complex"Acta Cryst.. D59. 185-187 (2003)

[文献書誌] Tachirov, T.H.: "Structure of a T7 RNA polymerase elongation complex at 2.9 A resolution"Nature. 420. 43-50 (2002)

[文献書誌] Sakamoto, K.: "Site-specific incorporation of an unnatural amino acid into proteins in mammalian cells"Nucleic Acids Res. 30. 4692-4699 (2002)

[文献書誌] Vassylyeva, M.N.: "Purification, crystallization and initial crystallographic analysis of RNA polymerase holoenzyme from Thermus thermophilus"Acta Cryst.. D58. 1497-1500 (2002)

[文献書誌] Kiga, D.: "An engineered Escherichia coli tyrosyl-tRNA synthetase for site-specific incorporation of an unnatural amino acid into proteins in eukaryotic translation and its application in a wheat germ cell-free system"Proc. Natl. Acad. Sci. U.S.A.. 99. 9715-9720 (2002)

[文献書誌] Nureki, O.: "an enzyme with a deep trefoil knot for the active-site architecture"Acta Cryst.. D58. 1129-1137 (2002)

[文献書誌] Vassylyev, D.G.: "Crystal structure of a bacterial RNA polymerase holoenzyme at 2.6 A resolution"Nature. 417. 712-719 (2002)

[文献書誌] Kitamura, A.: "Solution structure of an RNA fragment with the P7/P9.0 region and the 3'-terminal guanosine of the tetrahymena group I intron"RNA. 8. 440-451 (2002)

[文献書誌] Terada, T.: "Functional convergence of two lysyl-tRNA synthetases with unrelated topologies"Nature Struct. Biol.. 9. 257-262 (2002)

[文献書誌] Kitabatake, M.: "Indolmycin resistance of Streptomyces coelicolor A3(2) by induced expression of one of its two tryptophanyl-tRNA synthetases"J. Biol. Chem.. 277. 23882-23887 (2002)

[文献書誌] Hendrickson, T.L.: "Mutational separation of two pathways for editing by a class I tRNA synthetase"Mol. Cell. 9. 353-362 (2002)

[文献書誌] Hirao, I.: "An unnatural base pair for incorporating amino acid analogs into proteins"Nature Biotechnol.. 20. 177-182 (2002)

[文献書誌] Takasu, A.: "Analysis of Relative Positions of Ribonucleotide Bases in a Crystal Structure of Ribosome"Nucleotides and Nucleic Acids. 21. 449-462 (2002)

[文献書誌] Takasu, A.: "Classification of RNA Structures Based on Hydrogen Bond and Base-Base Stacking Patterns : Application for NMR structures"J. Biochem.. 132. 211-215 (2002)

[文献書誌] Sakamoto, T: "Solution structure of a SRP19 binding domain in human SRP RNA"J. Biochem.. 132. 177-182 (2002)

2002 年度実績報告書

横山茂之東京大学, 大学院・理学系研究科, 教授 (00159229)