2003 Fiscal Year Annual Research Report

RNAタンパク質複合体構造

Research Project

Project/Area Number	14035205
Research Institution	The University of Tokyo
Principal Investigator	横山茂之東京大学, 大学院・理学系研究科, 教授 (00159229)
Co-Investigator(Kenkyū-buntansha)	河合剛太千葉工業大学, 工学部, 助教授 (70211860)
Keywords	X線結晶構造解析 / NMR / 立体構造 / RNA / RNA結合タンパク質 / アミノアシルtRNA合成酵素 / RNAポリメラーゼ / リボゾーム
Research Abstract	1.X線結晶構造解析法:(1)古細菌のTyrRSとtRNA(Tyr)の複合体の結晶構造を決定し,古細菌と真核生物のTyrRSとtRNA(Tyr)が真正細菌のそれとは反応しない理由を明らかにした.(2)また,lleRSのCP1ドメインの結晶構造を決定した結果,間違って形成されたVal-tRNA(lle)を加水分解するメカニズムについて重要な知見を得ることができた.(3)古細菌のtRNAグアニントランスグリコシダーゼとtRNAとの複合体の結晶構造を決定し,tRNAがL字型とは異なるλ型の構造に変化して結合していることを明らかにした.(4)また,tRNAの3'末端のプロセシングに関与するRNase PHについてその結晶構造を決定し,tRNAとの結合モデルからその機能発現のメカニズムを示唆することができた.(5)古細菌のCCA付加酵素の結晶構造を決定した結果,真核生物および真正細菌のCCA付加酵素とは,アミノ酸配列のホモロジーが低いにも関わらず,触媒残基と基質であるCTPおよびATPとの位置関係がよく保存されていることがわかった(6)さらに,tRNAの連結反応に関与する高度好熱菌2'-5'RNAリガーゼの結晶構造を決定し,反応メカニズムについて考察した.(7)伸長反応中のT7 RNAポリメラーゼと基質アナログとの複合体の結晶構造を決定し,正しい基質を選別するメカニズムを明らかにした. 2.NMR法:(8)RNA分子の立体構造をその特徴によって分類するシステムを開発し,さらに,NMR法による構造解析結果を詳細に解析する手法として改良した.

Research Products

(13 results)

All Other

All Publications (13 results)

[Publications] Temiakov, D.: "Structural Basis for Substrate Selection by T7 RNA Polymerase"Cell. 116. 381-391 (2004)
[Publications] Kise, Y.: "A short peptide insertion crucial for angiostatic activity of human tryptophanyl-tRNA synthetase"Nat.Struct.Mol.Biol.. 11. 149-156 (2004)
[Publications] Sengoku, T.: "Crystallization and preliminary X-ray analysis of the helicase domains of Vasa complexed with RNA and an ATP analogue"Acta Crystallogr.D Biol Crystallogr.. 60. 320-322 (2004)
[Publications] Hirao, I.: "In vitro selection of RNA aptamers that bind to colicin E3 and structurally resemble the decoding site of 16S ribosomal RNA"Biochem.. 279. 8396-8402 (2003)
[Publications] Okabe, M.: "Divergent evolutions of trinucleotide polymerization revealed by an archaeal CCA-adding enzyme structure"EMBO J.. 22. 5918-5927 (2003)
[Publications] Polycarpo, C.: "Activation of the Pyrrolysine Suppressor tRNA Requires Formation of a Ternary Complex with Class I and Class II Lysyl-tRNA Synthetases"Mol.Cell. 12. 287-294 (2003)
[Publications] Kato, M.: "Crystal structure of the 2'-5'RNA ligase from Thermus thermophilus HB8"J.Mol.Biol.. 329. 903-911 (2003)
[Publications] Kobayashi, T.: "Structural basis for orthogonal tRNA specificities of tyrosyl-tRNA synthetases for genetic code expansion"Nat.Struct.Biol.. 10. 425-432 (2003)
[Publications] Ishii, R.: "Crystal structure of the tRNA processing enzyme RNase PH from Aquifex aeolicus"J.Biol.Chem.. 278. 32397-32404 (2003)
[Publications] Ishitani, R.: "by a posttr Alternative tertiary structure of tRNA for recognition anscriptional modification enzyme"Cel. 113. 383-394 (2003)
[Publications] Kaminishi, T.: "Crystallization and preliminary X-ray diffraction analysis of ribosomal protein L11 methyltransferase from Thermus thermophilus HB8"Acta Crystallogr.D Biol Crystallogr.. 59. 930-932 (2003)
[Publications] Baba, S.: "Role of the Zinc Fingers of HIV-1 Nucleocapsid Protein for Maturation of Genomic RNA"J.Biochem.. 134. 637-639 (2004)
[Publications] Someya, T.: "Analysis of local convergence in NMR structure calculation for RNA by a classification system for nucleic acid structure, CSNA"Nucleosides, Nucleotides and Nucleic Acids. (in press). (2004)

2003 Fiscal Year Annual Research Report

RNAタンパク質複合体構造

Principal Investigator

横山 茂之 東京大学, 大学院・理学系研究科, 教授 (00159229)

Research Products

[Publications] Temiakov, D.: "Structural Basis for Substrate Selection by T7 RNA Polymerase"Cell. 116. 381-391 (2004)

[Publications] Kise, Y.: "A short peptide insertion crucial for angiostatic activity of human tryptophanyl-tRNA synthetase"Nat.Struct.Mol.Biol.. 11. 149-156 (2004)

[Publications] Sengoku, T.: "Crystallization and preliminary X-ray analysis of the helicase domains of Vasa complexed with RNA and an ATP analogue"Acta Crystallogr.D Biol Crystallogr.. 60. 320-322 (2004)

[Publications] Hirao, I.: "In vitro selection of RNA aptamers that bind to colicin E3 and structurally resemble the decoding site of 16S ribosomal RNA"Biochem.. 279. 8396-8402 (2003)

[Publications] Okabe, M.: "Divergent evolutions of trinucleotide polymerization revealed by an archaeal CCA-adding enzyme structure"EMBO J.. 22. 5918-5927 (2003)

[Publications] Polycarpo, C.: "Activation of the Pyrrolysine Suppressor tRNA Requires Formation of a Ternary Complex with Class I and Class II Lysyl-tRNA Synthetases"Mol.Cell. 12. 287-294 (2003)

[Publications] Kato, M.: "Crystal structure of the 2'-5'RNA ligase from Thermus thermophilus HB8"J.Mol.Biol.. 329. 903-911 (2003)

[Publications] Kobayashi, T.: "Structural basis for orthogonal tRNA specificities of tyrosyl-tRNA synthetases for genetic code expansion"Nat.Struct.Biol.. 10. 425-432 (2003)

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

[Publications] Ishitani, R.: "by a posttr Alternative tertiary structure of tRNA for recognition anscriptional modification enzyme"Cel. 113. 383-394 (2003)

[Publications] Kaminishi, T.: "Crystallization and preliminary X-ray diffraction analysis of ribosomal protein L11 methyltransferase from Thermus thermophilus HB8"Acta Crystallogr.D Biol Crystallogr.. 59. 930-932 (2003)

[Publications] Baba, S.: "Role of the Zinc Fingers of HIV-1 Nucleocapsid Protein for Maturation of Genomic RNA"J.Biochem.. 134. 637-639 (2004)

[Publications] Someya, T.: "Analysis of local convergence in NMR structure calculation for RNA by a classification system for nucleic acid structure, CSNA"Nucleosides, Nucleotides and Nucleic Acids. (in press). (2004)

横山茂之東京大学, 大学院・理学系研究科, 教授 (00159229)