2001 Fiscal Year Final Research Report Summary
Elucidation of the construction principle of the mitochondria translation system by means of structural biology
| Project/Area Number |
11308024
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Structural biochemistry
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| Research Institution | The University of Tokyo |
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Principal Investigator |
WATANABE Kimitsuna The University of Tokyo, Graduate school of Frontier Sciences, Professor, 大学院・新領域創成科学研究科, 教授 (00134502)
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| Co-Investigator(Kenkyū-buntansha) |
OHTSUKI Takashi The University of Tokyo, Graduate school of Engineering, Research associate, 大学院・工学系研究科, 助手 (80321735)
SUZUKI Tsutomu The University of Tokyo, Graduate school of Frontier Sciences, Lecturer, 大学院・新領域創成科学研究科, 講師 (20292782)
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| Project Period (FY) |
1999 – 2001
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| Keywords | Translation System / mitochondria / EF-Tu / tRNA / Seryl-tRNA synthetase / Nematode / ribosome / X-ray analysis |
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| Research Abstract |
The aim of this research is to elucidate the construction principle of the mitochondria! translation system, by analyzing the functional structures of each component of the animal mitochondrial translation system using X-ray analysis and NMR spectroscopy, focusing on the functional characteristics and structural abnormalities of the components. The following results have been obtained for these three years.
(1) Bovine mitochondrial in vitro translation system has been established by optimizing concentration of each component and reaction conditions, which has similar efficiency to that of 」. co// system in terms of poly(U)-dependent poly(Phe) synthesis.
(2) Ribosomes are exchangeable between E. coli and mitochondrial systems, as far as tRNA and translation factors of the same origins are used, and EF-Tu of E. coli origin is used. Thus, the ribosomes are functionally equivalent between E. coli and mitochondrial systems.
(3) It was verified that 2 species of serine tRNAs with unusual secondary structures can be functional. The low efficiency of the D arm-lacking serine tRNA is derived from a defect in its binding to the ribosomal A site.
(4) These two serine tRNAs were recognized by a single seryl-tRNA synthetase (serRS) and its recognition sites on the two tRNAs were identified.
(5) A unique EF-Tu was found which recognized almost all the T arm-lacking tRNAs in nematode mitochondria. It had 57 amino acid extension at the C terminus which was considered to complement the T arm-lacking part of tRNAs. Another EF-Tu recognizing two D arm-lacking serine tRNAs was specific for serine.
(6) Crystallization of these translation factors are being under way. We have obtained a needle-like srystal for SerRS. The X-ray analysis will be made in future.
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Research Products
(15 results)
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[Publications] Yokogawa, T., Shimada, N., Takeuchi, N., Ueda, T., Nishikawa, K., Spremulli, L., and Watanabe K.: "Characterization and tRNA recognition of mammalian mitochondrial Seryl tRNA synthetase"J. Biol. Chem.. 275. 19913-19920 (2000)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Suzuki, T., Terasaki, M., Takemoto-Hori, C., Hanada, T., Ueda, T., Wada, A., and Watanabe, K.: "Structural compensation for deficit of rRNA with proteins in mammalian mitochondrial ribosome; systematic analysis of protein components of the large ribosomal subunit from mammalian mitochondria"J. Biol. Chem.. 276. 21724-21736 (2001)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Suzuki, T., Terasaki, M., Takemoto-Hori, C., Hanada, T., Ueda, T., Wada, A. and Watanabe, K.: "Proteomic Analysis of the Mammalian Mitochondrial Ribosome; Identification of protein components in the 28S Small Subunit"J. Biol. Chem.. 276. 33181-33195 (2001)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Hanada, T., Suzuki, T., Yokogawa, T., Takemoto-Hori, C., Sprinzl, M. and Watanabe, K.: "Translation ability of mitochondrial tRNAs^<Ser> with unusual secondary structures in an in vitro translation system of bovine mitochondria"Genes Cells. 6. 1019-1030 (2001)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Ohtsuki, T., Watanabe, Y., Takemoto, C., Kawai, G., Ueda, T., Kita, K., Kojima, S., Kaziro, Y., Nyborg, J and Watanabe, K.: "An "Elongated" Translation Elongation Factor Tu for Truncated tRNAs in Nematode Mitochondria"J. Biol. Chem.. 276. 21571-21577 (2001)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Takeuchi, N., Vial, L., Panvert, M., Schmidt, E., Watanabe K., mechulam Y. and Blanquet, S.: "Recognition of tRNAs by Methionyl-tRNA Transformylase from mammalian Mitochondria"J. Biol. Chem.. 276. 20064-20068 (2001)
Description
「研究成果報告書概要(欧文)」より
