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Functional analysis of chloroplast genes by chloroplast transformation

Research Project

Project/Area Number 10440238
Research Category

Grant-in-Aid for Scientific Research (B).

Allocation TypeSingle-year Grants
Section一般
Research Field 植物生理
Research InstitutionNagova University

Principal Investigator

SUGITA Mamoru  Nagoya University, Human Informatics, Professor, 人間情報学研究科, 教授 (70154474)

Co-Investigator(Kenkyū-buntansha) SUGIURA Masahiro  Nagoya City University, Graduate School of Natural Sciences, Professor, 大学院・システム自然科学研究科, 教授 (80027044)
廣瀬 哲郎  名古屋大学, 遺伝子実験施設, 助手 (30273220)
Project Period (FY) 1998 – 2000
Project Status Completed (Fiscal Year 2000)
Budget Amount *help
¥13,800,000 (Direct Cost: ¥13,800,000)
Fiscal Year 2000: ¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 1999: ¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 1998: ¥7,200,000 (Direct Cost: ¥7,200,000)
Keywordstobacco / cyanobacteria / chloroplast transformation / ycf / small stable RNA / knockout / ribonucleoprotein / phage-type RNA polymerase / 葉緑体 / ycf5 / 光合成 / DNAマイクロアレイ / 原色素体 / DNA
Research Abstract

This research purpose is to define the function of hypothetical chloroplast reading frames (ycf) and to understand the mechanism of regulation of chloroplast gene expression. The novel findings obtained are as follows.
1. Two targeted ycf5-disruptants were obtained. These disruptants were uncapable of electron-fow from photosystem II to photosystem I.This indicates that ycf5 is involved in electron transport in photosynthesis.
2. The gene encoding a small stable RNA (tmRNA) was isolated and characterized in the unicellular cyanobacterium, Synechococcus PCC6301. A tmRNA gene is not present in higher plant chloroplast DNA, suggesting that a ancestral chloroplast tmRNA gene lost during evolution.
3. By immunoprecipitation, gel filtration, and western blot analysis, we demonstrated that tobacco chloroplast ribonucleoproteins (cpRNPs) are abundant stromal proteins that exist as complexes with ribosome-free mRNAs. In addition, an in vitro mRNA degradation assay revealed that the cpRNPs act as stabilizing factors for nonribosome-bound mRNAs in the stroma.
4. We isolated and characterized the three genes, NsRpoT-A, NsRpoT-B and NsRpoT-C, encoding phage-type single subunit RNA polymerase from Nicotiana sylvestris. The gene product of NsRpoT-B was demonstrated to be imported into mitochondria and plastids. This provides a new insight of evolution of plant organelles.

Report

(4 results)
  • 2000 Annual Research Report   Final Research Report Summary
  • 1999 Annual Research Report
  • 1998 Annual Research Report
  • Research Products

    (32 results)

All Other

All Publications (32 results)

  • [Publications] Watanabe,T.,Sugita,M.,Sugiura,M.: "Identification of 10Sa RNA (tmRNA) homologues from the cyanobacterium Synechococcus sp. strain PCC63O1 and related organisms."Biochim.Biophys.Acta. 1396. 97-104 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Sugiura,M.,Hirose,T.,Sugita,M.: "Evolution and mechanism of translation in chloroplasts."Annu.Rev.Genet.. 32. 437-459 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Wakasugi,T.,Sugita,M.,Tsudzuki,T.,Sugiura,M.: "Updated gene map of tobacco chloroplast DNA."Plant Mol.Biol.Rep.. 16. 231-241 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Nakamura,T.,Ohta,M.,Sugiura,M.,Sugita,M.: "Chloroplast ribonucleoproteins are associated with both mRNAs and intron-containing precursor tRNAs."FEBS Lett.. 460. 437-441 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Hirose,T.,Kusumegi,T.,Tsudzuki,T.,Sugiura,M.: "RNA editing sites in tobacco chloroplast transcripts : editing as a possible regulator of chloroplast RNA polymerase activity."Mol.Gen.Genet.. 262. 462-467 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Nakamura,T.,Ohta,M.,Sugiura,M.,Sugita,M.: "Chloroplast ribonucleoproteins function as a stabilizing factor of ribosome-free mRNAs in the stroma."J.Biol.Chem.. 276. 147-152 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] 杉田護: "シリーズ・ニューバイオフィジックス2巻 遺伝子の構造生物学"(日本生物物理学会編)共立出版. 208 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] 杉田護: "朝倉植物生理学講座第3巻 光合成"朝倉書店(印刷中). (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Watanabe, T., Sugita, M., Sugiura, M.: "Identification of 10Sa RNA (tmRNA) homologues from the cyanobacterium Synechococcus sp.strain PCC6301 and related organisms."Biochim.Biophys.Acta. 1396. 97-104 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Sugiura, M., Hirose, T., Sugita, M.: "Evolution and mechanism of translation in chloroplasts."Annu.Rev.Genet.. 32. 437-459 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Wakasugi, T., Sugita, M., Tsudzuki, T., Sugiura, M.: "Updated gene map of tobacco chloroplast DNA."Plant Mol.Biol.Rep.. 16. 231-241 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Nakamura, T., Ohta, M., Sugiura, M., Sugita, M.: "Chloroplast ribonucleoproteins are associated with both mRNAs and intron-containing precursor tRNAs."FEBS Lett.. 460. 437-441 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Hirose, T., Kusumegi, T., Tsudzuki, T., Sugiura, M.: "RNA editing sites in tobacco chloroplast transcripts : editing as a possible regulator of chloroplast RNA polymerase activity."Mol.Gen.Genet.. 262. 462-467 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Nakamura, T., Ohta, M., Sugiura, M., Sugita, M.: "Chloroplast ribonucleoproteins function as a stabilizing factor of ribosome-free mRNAs in the stroma."J.Biol.Chem.. 276. 147-152 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Sugita,C.,Sugiura,M.,Sugita,M.: "A novel nucleic acid-bindingprotein in the cyanobacterium Synechococcus sp.PCC6301: a 33 kDa soluble polypeptide with high sequencesimilarity to ribosomal protein S1."Mol.Gen.Genet.. 263. 655-663 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Yukawa,Y.,Sugita,M.,Choisne,N.Small,I.D.,Sugiura,M.: "The TATA motif, the CAA motif and the poly (T) transcription termination motif are all important for transcription re-initiation on plant tRNA genes."The Plant J.. 22. 439-447 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Nakamura,T.,Ohta,M.,Sugiura,M.,Sugita,M.: "Chloroplast ribonucleoproteins function as a stabilizing factor of ribosome-free mRNAs in the stroma."J.Biol.Chem.. 276. 147-152 (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Hara,K.,Sugita,M.,Aoki,S.: "Cloning and characterization of the cDNA for a plastid sigma factor from the moss Physcomitrella patens."Biochim.Biophys.Acta. 1517. 302-306 (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] 杉田護: "色素体遺伝子の転写を支配する核ゲノム"ブレインテクノニュース. 79. 22-24 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 杉田護: "朝倉植物生理学講座 第3巻 光合成(佐藤公行編集)"朝倉書店(印刷中). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Sugita, C., Mutsuda, M., Sugiura, M., Sugita, M.: "Targeted deletion of genes for eukaryotic RNA-binding proteins, Rbp1 and Rbp2, in cyanobacterium Synechococcus sp. strain PCC7942: Rbp1 gene is indispensable for cell growth at low temperatures."FEMS Microbiol. Lett.. 176. 142-147 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Nakamura, T., Ohta, M., Sugiura, M., Sugita, M.: "Chloroplast ribonucleoproteins are associated with both mRNAs and intron-containing precursor tRNAs."FEBS Lett.. 460. 437-441 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Mutsuda, M., Sugiura, M., Sugita, M.: "Physiological characterization of RNA-binding protein-deficient cells from Synechococcus sp. strain PCC7942."Plant Cell Physiol.. 40. 1203-1209 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Kapoor, S., Sugiura, M.: "Identification of two essential sequence elements in the nonconcensus type II PatpB-290 plastid promoter by using plastid transcription extracts from cultured tobacco BY-2 cells."Plant Cell. 11. 1799-1810 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] 杉田 護、杉浦昌弘、広瀬哲郎: "遺伝子遺伝子発現における転写後調節"蛋白質 核酸 酵素. 45. 132-138 (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] Sugita, C., Sugiura, M., Sugita, M.: "A novel nucleic acid-binding protein in the cyanobacterium Synechococcus sp. PCC6301: a 33 kDa soluble polypeptide with high sequence similarity to ribosomal protein S1."Mol. Gen. Genet.. (印刷中). (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] 宮城 徹: "Transcript analysis of tobacco plastid rps2/atpI/H/F/A operon reveals the existence of a non-consensus type II(NCII)promoter upstream to atpI coding sequence." Mol.Gen.Genet.257. 299-307 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] 渡辺恒暁: "Identification of 10Sa RNA (tmRNA)homologues from the cyanobacterium Synechococcus sp.strain PCC6301 and related organisms." Biochim.Biophys.Acta. 1396. 97-104 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] 若杉達也: "Update gene map of tobacco chloroplast DNA." Plant Mol.Biol.Rep.16. 231-241 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] 杉浦昌弘: "Evolution and mechanism of translation in chloroplasts." Annu.Rev.Genet.32. 437-459 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] 廣瀬哲郎: "Translation of tobacco chloroplast rps14 mRNA depends on a Shine-Dalgarno-like sequence in the 5'-untranslated region but not on internal RNA edition in the coding region." FEBS Letters. 430. 257-260 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] 赤間一仁: "Plant cytosolic tRNA^<His> possesses an exceptional C_<54> in the canonical TΨC loop." Nucleic Acids Res.26. 2708-2714 (1998)

    • Related Report
      1998 Annual Research Report

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Published: 1998-04-01   Modified: 2016-04-21  

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