| Project/Area Number |
07044183
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| Research Category |
Grant-in-Aid for international Scientific Research
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| Allocation Type | Single-year Grants |
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| Section | Joint Research |
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| Research Institution | University of Tokyo |
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Principal Investigator |
WATANABE Kimitsuna University of Tokyo, Professor, 大学院・工学系研究科, 教授 (00134502)
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| Co-Investigator(Kenkyū-buntansha) |
NITTA Itaru University of Tokyo, 大学院・工学系研究科, 助手 (30272404)
UEDA Takuya University of Tokyo, 大学院・工学系研究科, 助教授 (80184927)
HARRY Noller カリフォルニア大学, RNA分子生物学センター, 教授
KNUD Nierhau マックスープランク分子遺伝学研究所, 教授
NIERHAUS H.Knud Max-Planck Institute
NOLLER F.Harry University of California
NOLLER F.Har カリフォルニア大学, RNA分子生物学センター, 教授
NIERHAUS H.K マックスープランク分子遺伝学研究所, 教授
HIERHAUS Knu マックスプランク分子遺伝学研究所, 教授
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥5,200,000 (Direct Cost: ¥5,200,000)
Fiscal Year 1996: ¥2,800,000 (Direct Cost: ¥2,800,000)
Fiscal Year 1995: ¥2,400,000 (Direct Cost: ¥2,400,000)
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| Keywords | Pyridine / Tertiary amine / ribosome / rRNA / peptidyl transterase / translation / RIP / 生体外蛋白質合成系 / リボソームサブユニット / リボソーム蛋白質 / リボソームRNA / リボソーム再編成 / ピリジン |
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| Research Abstract |
We found that a high concentration (40-60%) of pyridine, an aromatic tertiary amine catalyst, is able to promote the translation on ribosomes without soluble protein factors of chemical energy sources. This novel translation system was called the pyridine system, which could produce oligophyenylalanine with chain lengths of up to decamer and polylysine with chain lengths of around 40mer, depending on the corresponding templates, poly (U) and poly (A), respectively. In poly (UC) -dependent oligo (serine-leucine) synthesis, oligopeptides with a serine and leucine alternate sequence were the main products. The template dependency is most prominent at 60% pyridine, but not observed at 40% or 50% pyridine. The optimal temperature range of the reaction was rather wide between 30 and 60ﾟC,which may reflect no requirement for temperature-sensitive soluble protein factors for the reaction. The ranges of the optimal concentrations of K^+ (200-500 mM) and Mg^<2+> (1-50 mM) in the pyridine system
… More
are also rather wide, quite different from those in the usual aqueous translation system.
Current investigations have suggested that rRNAs play a central role in the ribosomal functions, although the conclusive evidence has not been provided ; some papers have reported that the eubacterial peptidyltransferation, the peptide-bond formation on the ribosomal large subunits, scarcely requires their proteins. Here we demonstrate that E.coli 23S rRNAs either extracted from ribosomes or synthesized in vitro are able to promote the peptide-bond formation (phenylalanylphenylalanine synthesis from phenylalanyl-tRNA^<Phe>) without template in the presence of 40% pyridine. The reaction required 23S rRNAs or the transcripts in the folded state, and was inhibited with chloramphenicol but not with cycloheximide as well as by site-directed digestion with RNase H of 23S rRNA hybridized with some complementary oligodeoxyribonucleotides. These results strongly suggest that 23S rRNA is directly involved in the peptide bond formation. Less
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