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1996 Fiscal Year Final Research Report Summary

Molecular and cellular studies on endonuclear symbionts

Research Project

Project/Area Number 07044202
Research Category

Grant-in-Aid for international Scientific Research

Allocation TypeSingle-year Grants
SectionJoint Research
Research InstitutionYamaguchi University

Principal Investigator

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

Co-Investigator(Kenkyū-buntansha) FOKIN Sergei L  St.-Petersburg State University, Senior Researcher, 生物学部, 研究員
SKOBLO Inna I  St.-Petersburg State University, Senior Researcher, 生物学部, 研究員
RAUTIAN Maria S  St.-Petersburg State University, Senior Researcher, 生物学部, 研究員
OSSIPOV Dmitry V  St.-Petersburg State University, Professor, 生物学部, 教授
BRIGGE Theodor  Stuttgart University, Assistant, 動物学部, 助手
GORTZ Hans-Dieter  Stuttgart University, Professor, 動物学部, 教授
ISHIKAWA Hajime  Graduate School of Tokyo University, Professor, 大学院・理学系研究科, 教授 (70012482)
HORI Manabu  Yamaguchi University, Assistant, 理学部, 助手 (00253138)
Project Period (FY) 1995 – 1996
KeywordsParamecium / Holospora / Proton pomp / Endosymbiotic bacterium / Endonuclear symbiotic bacterium / Protozoa / Ciliates / Infection
Research Abstract

1.Acidification of inside pH of Paramecium digestive vacuole is induced by a proton pomp (V-ATPase) of the vacuole membrane, and the acidification is indispensble phenomenon for Holospora's escape from the vacuole.
2.Reproductive forms of Holospora obtusa promotes the host cell division not only in low temperature (4゚C) but also in 25゚C.
3.When the reproductive forms differentiate to the infectious long forms, large amounts of cell-wall antigen (IR-4-1 antigen) are released, or the infectious forms are continuously shedding the antigens, and the shedded antigens stick to the macronuclear envelope and make the envelope unable to pass large molecules such as immunoglobulins inside the macronuclear envelope. This may be a cause for the host death when the macronucleus is filled with the infectious long forms. A mutant strain without the IR-4-1 antigen could not induce the host death even if the host macronucleus was filled with the infectious forms.
4.Genes of stress proteins, groES and groEL homolog, were cloned in H.obtusa.
5.Infectious form-specific 4.5 K protein of H.obtusa was purified and aminoacid sequences was clarified in the total length, and also the gene was cloned. Immunoelectron microscopy with a polyclonal antibody showed that the antigen was localized in the periplasm. Northern blot analysis using the cDNA showed that transcriptio was carried out in intermediate form of the bacterium which appeared in differentiation from the reproductive to the infectious form.
6.Indirect immunofluorescence microscopy and immunoblot with antibodies against Paramecium caudatum actin showed that H.obtusa had a protein with an epitope for the antibody.
Using pulse field gel electrophoresis (PFGE), is was found that genome sizes of H.undulata, H.obtusa and H.recta were 1800,1740 and 1800 kbp respectively.

  • Research Products

    (18 results)

All Other

All Publications (18 results)

  • [Publications] Miki Kawai: "Partial characterization and cryopreservation of the isolated symbiotic bacterium Holospora recta of the ciliate Paramecium caudatum." Zoological Science. 13. 679-684 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Masahiro Fujishima: "Macronuclear nucleoli specific protein of Paramecium caudatum." European Journal of Protistology. 32. 25-31 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Masahiro Fujishima: "Qantitative changes in periplasmic proteins of the macronuclear-specific bacterium Holospora obtusa in the infection process of the ciliate Paramecium caudatum." Journal of Eukaryotic Microbiology. (Accepted). (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Maki Murata-Hori: "Releases substances from Tetrahymena thermophila arrest the cell cycle at G1 phase and removal of the substances induces highly synchronized cell division." European Journal of Protistology. 32. 481-490 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hideo Dohra: "Cloning and sequencing of a periplasmic 54 kDa peptide gene of the macronucleus-specific symbiont Holospora obtusa of the ciliate Paramecium caudatum." Zoological Science. (In press). (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sergei I.Fokin: "Holospora species infecting the nuclei of Paramecium appear to belong into two groups of bacteria." European Journal of Protistology. 32. 19-24 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Maria Rautian: "Transformation of a Paramecium caudatum cloneresistant to infection by intranuclear symbiotic bacteria of the genus Holospora" European Journal of Protistology. 32. 135-140 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Anna.S.Timofeeva: "The determination of genome size of symbiotic bacteria Holospora undulata by pulse-field gel electrophoresis (PFGE)." Tsitologia. (In press). (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 藤島政博: "総説 ゾウリムシとホロスポラの共生における宿主-共生生物間相互作用" 原生動物学雑誌. (印刷中). (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Miki Kawai: "Partial characterization and cryopreservation of the isolated symbiotic bacterium Holospora recta of the ciliate Paramecium caudatum." Zoological Science. 13. 679-684 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Masahiro Fujishima: "Macronuclear nucleoli spesific protein of Paramecium caudatum." European Journal of Protistology. 32. 25-31 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Masahiro Fujishima: "Qantitative changes in periplasmic proteins of the macronuclear-specifi bacterium Holospora obtusa in the infection process of the ciliate Paramecium caudatum." Journal of Eukaryotic Microbiology. Accepted. (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Maki Murata-Hori: "Releases substances from Tetrahymena thermophila arrest the cell cyc at G1 phase and removal of the substances induces highly synchronize cell division." European Journal of Protistology. 32. 481-490 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hideo Dohra: "Cloning and sequencing of a periplasmic 5.4 kDa peptide gene of the macronucleus-specific symbiont Holospora obtusa of the ciliate Paramecium caudatum." In press. (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sergei I.Fokin: "Cloning and sequencing of a periplasmic 5.4 kDa peptide gene of the macronucleus-specific symbiont Holospora obtusa of the ciliate Paramecium caudatum." Zoological Science. 32. 19-24 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Maria Rautian: "Transformation of a Paramecium caudatum cloneresistant to infection by intranuclear symbiotic bacteria of the genus Holospora" European Journal of Protistology. 32. 135-140 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Anna.S.Timofeeva: "The determination of genome size of symbiotic bacteria Holospora undulata by pulse-field gel electrophoresis (PFGE)." Tsitologia. In press. (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Masahiro Fujishima: "Review host-symbiont interactions in Paramecium-Holospora symbiosis" Japan Journal of Protozoology. In press. (1997)

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

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Published: 1999-03-09  

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