2002 Fiscal Year Final Research Report Summary
Chloroplast RNA-binding proteins involved in RNA processing and translation
| Project/Area Number |
12440226
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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 |
植物生理
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| Research Institution | Nagoya City University |
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Principal Investigator |
MASAHIRO Sugiura Nagoya City Univ., Graduate School of Natural Sciences, Prof., 大学院・システム自然科学研究科, 教授 (80027044)
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| Co-Investigator(Kenkyū-buntansha) |
SAKURAI Norihiko Nagoya City Univ., Graduate School of Natural Sciences, Assistant Prof., 大学院・システム自然科学研究科, 講師 (00255233)
MORIYAMA Akihiko Nagoya City Univ., Graduate School of Natural Sciences, Prof., 大学院・システム自然科学研究科, 教授 (50145744)
EIICHI Tanimoto Nagoya City Univ., Graduate School of Natural Sciences, Prof., 大学院・システム自然科学研究科, 教授 (90080283)
OBOKATA Junichi Nagoya City Univ., Gene Research Center, Associate Prof., 遺伝子実験施設, 助教授 (50185667)
TSUDZUKI Takahiko Aichi-Gakuin Univ., Dept.of Information & Policy Studies, Prof., 情報社会政策学部, 教授 (60064896)
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| Project Period (FY) |
2000 – 2002
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| Keywords | chloroplast / tobacco / cis-element / translation / RNA editing / mRNA / Shine-Dalgarno / in vitro system |
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| Research Abstract |
(1)Conditions for chloroplast preparation
As pea is suitable for biochemical studies, we optimized its growth conditions, leaf harvest and preparation of isolated of isolated chloroplasts. Then, we searched for RNA editing sites in pea chloroplast transcripts.
(2)Analysis of cis-and trans-factors for chloroplast translation
Thirty mRNAs from tobacco chloroplasts have no Shine-Dalgarno-like sequences in their 5'-UTRs and require trans-factors for translation. Using our chloroplast in vitro translation system, we identified cis-elements for translation. Using our chloroplast in vitro translation system, we identified cis-elements for translation of chloroplast atpB mRNA. The tobacco chloroplast rps2 mRNA possesses a Shine-Dalgarno-like sequence, but surprisingly this sequence was found to be inhibitory for translation.
(3)Analysis of factors involved in chloroplast RNA editing
To analyze the molecular mechanism of RNA editing in chloroplasts, we developed for the first time a chloroplast in vitro system. Using this system, we identified site-recognition factors for psbL mRNA and psbE mRNA, which are 25 kDa and 56 kDa proteins, respectively. We tried to isolate these proteins but so far unsuccessful.
(4)RNA editing analysis in pea chloroplasts
As a second model system, we developed a chloroplast in vitro system from isolated pea chloroplasts. Using this system, we found no editing activity for psbE mRNA as pea lost its editing site. This indicates that editing sites and editing activities are co-evolued.
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Research Products
(12 results)
