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2000 Fiscal Year Final Research Report Summary

Interaction Between Endosymbiont Holospora and the Cell

Research Project

Project/Area Number 11694211
Research Category

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

Allocation TypeSingle-year Grants
Section一般
Research Field 動物生理・代謝
Research InstitutionYamaguchi University

Principal Investigator

FUJISHIMA Masahiro  Yamaguchi University, Faculty of Science, Professor, 理学部, 教授 (40127783)

Co-Investigator(Kenkyū-buntansha) HORI Manabu  Yamaguchi University, Faculty of Science, Assistant, 理学部, 助手 (00253138)
Project Period (FY) 1999 – 2000
KeywordsEndonuclear symbiont / Holospora / Stress responces / Paramecium / Endosymbiosis / Gene expressions / Targeting / Nuclear membrane
Research Abstract

1. We found that nuclear envelope-recognition sensor substances of H.obtusa and H.undulata were lipo-polysaccharides (LPSs) on their outer membranes. Indirect immunofluorescence microscopy and immuno-electron microscopy with monoclonal antibodies for these LPSs showed that the antibodies reacted with their target nuclear envelopes.
2. Holospora penetrates the target nuclear envelope with its specific tip. When this special tip was cut off from about 3,400 infectious forrns with a micro-dissection system and loaded on SDS-PAGE, 2 bands appeared with silver staining. These area of SDS-PAGE gels were cut and inside materials were electroeluted. Using these eluted substances as antigens, we succeeded to develop a monoclonal antibody specific for the special substance with indirect immunofluorescence microscopy.
3. Eighteen spots were cut off from 2D-SDS-PAGE of H.obtusa and the micro-sequences of the purified proteins from their N-terminals were clarified and homology checks with known proteins were carried out. Also, their ORFs were searched from H.obtusa genomic DNA sequences so far clarified. As a the results, only 3 ORFs were found among 18 proteins, and 8 proteins showed high homologies with known proteins. The fact that only 3 ORF were detected shows that majority of the H.obtusa proteins may derived from the host cells.
4. We found 12 host genes that were depressed and 3 host genes by the infection of H.obtusa by differential display (DD) method. Partial DNA sequences of 4 genes of the former ones were clarified. One of the 4 genes showed was an immobilization antigen A homologous gene of Paramecium tetraurelia.

  • Research Products

    (13 results)

All Other

All Publications (13 results)

  • [Publications] Hideo Dohra: "Cell structure of the infectious form of Holospora an endonuclear symbiotic bacterium of the ciliate Paramecium."Zoological Science. 16. 93-98 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Eriko Osaki: "Purification of germinal-vesicle-breakdown-inducing proteins from Tetrahymena pyriformis."European Journal of Protistology. 36. 459-464 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hideo Dohra: "Effects of antibiotics on early infection process of a macronuclear endosymbiotic bacterium Holospora obtusa of Paramecium caudatum"FEMS Microbiology Leteter. 179. 473-477 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Miki Kawai: "Invasion of the macronucleus of Paramecium caudatum by the bacterium Holospora obtusa Fates of the bacteria and timings of invasion steps."European Journal of Protistology. 36. 46-52 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ilys N.Skovorodkin: "Fates of the endonuclear symbiotic bactena Holospora obtusa and Holospora undulata injected into the macronucleus of Paramecium caudatum"European Journal of Protistology. (in press). (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Manabu Sugii: "Purification of GVBD-inducing protein from the ciliate Tetrahymena thermophila."European Journal of Protistology. (in press). (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 藤島政博: "細胞内共生細胞とその遺伝.(ゾウリムシの遺伝学,樋渡宏一 編集)"東北大学出版会. 14 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hideo Dohra and Masahiro Fujishima: "Cell structure of the infectious form of Holospora an endonuclear symbiotic bacterium of the ciliatearamecium"Zoological Scienc. 16. 93-98 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Eriko Osakii and Masahiro Fujishima: "Purification of Germinal-Vesicle-Breakdown-inducing protein from the ciliate Tetrahymenapyrifomis"European Journal of Protistology. 36. 459-464 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hideo Dohra and Masahiro Fujishima: "Effects of antibiotics on early infection process of a macronuclear endosymbiotic bacterium Holospora obtusa of Paramecium caudatum"FEMS Microbiology Leteter. 179. 473-477 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Miki Kawai and Masahiro Fujishima: "Invasion of the macronucleus of Paramecium caudatum by the bacterium Holospora obtusa Fates of the bacteria and timings of invasion steps."European Journal of Protistology. 36. 46-52 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ilya N.Skovorodkin, Sergei I.Fokin and Masahiro Fujishima: "Fates of the endonuclear symbiotic bacteria Holospora obtusa and Holospora undulatan jected into the macronucleus of Paramecium caudatum"European Journal of Protistology. 36. 46-52 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Manabu Sugii and Masahiro Fujishima: "Purification of GVBD-inducing protein from the ciliate Tetrahymena thermophila."Journal of Eukaryotic Microbiology. (in press). (2001)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2002-03-26  

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