| Project/Area Number |
11480198
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Molecular biology
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| Research Institution | THE UNIVERSITY OF TOKYO |
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Principal Investigator |
UEDA Takuya GRADUATE SCHOOL OF FRONTIER SCIENCES, THE UNIVERSITY OF TOKYO, PROFESSOR, 大学院・新領域創成科学研究科, 教授 (80184927)
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| Co-Investigator(Kenkyū-buntansha) |
SUZUKI Tsutomu GRADUATE SCHOOL OF FRONTIER SCIENCES, THE UNIVERSITY OF TOKYO, LECTURER, 大学院・新領域創成科学研究科, 講師 (20292782)
OHYA Yoshikazu GRADUATE SCHOOL OF FRONTIER SCIENCES, THE UNIVERSITY OF TOKYO, PROFESSOR, 大学院・新領域創成科学研究科, 教授 (20183767)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥14,300,000 (Direct Cost: ¥14,300,000)
Fiscal Year 2000: ¥4,200,000 (Direct Cost: ¥4,200,000)
Fiscal Year 1999: ¥10,100,000 (Direct Cost: ¥10,100,000)
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| Keywords | tRNA / aminoacylation / methylation / transcription / polysemous codon / codon recognition / 翻訳 / Candida / メチル化 |
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| Research Abstract |
We demonstrated that codon CUG is assigned simultaneously both-as serine arid leucine codons in several- Candida-species, using analyzes of tRNA-and genetic techniques. In order to evaluate biological significance of the polysemous decoding in Candid strains, we are addressing quantitative manipulation of leucine acceptance of responsible tRNA. We already showed that leucine acceptance is governed by the presence of 1-methylguanosine at the position 37 adjacent to the anticodon. Moreover it is suggested that A at 73 position is also significantly involved in leucylation. In this project/we prepared various transcripts with mutation and modification enzyme for m1G from E. coli by overexpression method. Furthermore, seryl-tRNA and leucyl-tRNA synthetases were also expressed in E. coli and purified. By determination of enzymatic parameters of leucylation reaction using mutants, it appeared that methyl group at position 37 is indispensable for leucine *. Acceptance. Furthermore, mutation G to A at position 73 increased Kcat/Km values 400-fold. Base upon these observations, we will introduce these mutants genes into Candida cells and alternation of cell function will be evaluated.
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