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Physiological studies on the function of double-stranded ribonuclease during the larval-pupal metamorphosis of the sklkworm, Bombyx mori

Research Project

Project/Area Number 04454066
Research Category

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

Allocation TypeSingle-year Grants
Research Field 蚕糸学
Research InstitutionKyoto Institute of Technology

Principal Investigator

FURUSAWA Toshiharu  Kyoto Inst. Technol., Dep. Applied Biol., Professor, 繊維学部, 教授 (70127166)

Co-Investigator(Kenkyū-buntansha) MORI Hajime  Kyoto Inst. Technol., Dep. Applied Biol., Assist. Prof., 繊維学部, 助手 (80201812)
Project Period (FY) 1992 – 1993
Project Status Completed (Fiscal Year 1993)
Budget Amount *help
¥6,600,000 (Direct Cost: ¥6,600,000)
Fiscal Year 1993: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1992: ¥5,400,000 (Direct Cost: ¥5,400,000)
KeywordsMetamorphosis of insects / Chromatin droplets / DNA fragments / Alimentary canal / Apoptosis / DNase / Double-stranded ribonuclease / Endonuclease / カイコ / 細胞質多角体病ウイルス / 変態
Research Abstract

A double-stranded ribonuclease (dsRNase), which exists in the midgut epithelium and the digestive juice of the silkworm, Bombyx mori, was partially purified by using DEAE, CM and G-75 column chromatography. On the elution profile of nuclease activity of each chromatogram, the elution region of dsRNase activity was coincided with those of DNase and ssRNase activities. In order to isolate the dsRNase from DNase and single-stranded RNase (ssRNase), dsRNase activity fraction on CM colum chromatography was applied to native DNA-cellulose column chromatography, and then eluted by the stepwaise method of the increasing of NaCl concentration. The dsRNase was still eluted with DNase and ssRNase from 0.25 through 0.5 M NaCl. Further investigation clarify that both DNase and dsRNase was eluted from 0.25 through 0.3 M NaCl, and that ssRNase with the DNase and dsRNase from 0.4 through 0.5 M NaCl. The activities of DNase and dsRNase, which were obtained from nuclease activity fraction on DNA cellulo … More se colum chromaography, were inhibited with the increasing of single-stranded RNA (ssRNA) content, and degraded a circular DNA (pUC 119). These results suggest that an endonuclease, which will be capable of attacking DNA, dsRNA and ssRNA, exists in the digestive juice of B.mori.
The endonuclease indicates a high activity in the midgut and the digestive juice during larval-pupal metamorphosis. At this period, chromatin droplets were observed in the content of the inside of the midgut, and also an analysis of DNA extracted from the content exhibited a ladder-like electrophoretic pattern on an agarose gel. On the basis of these data, an experiment was undertaken to investigate whether the endonuclease in the digestive juice will be concerned with the fragmentation of DNA.The in vitro incubation of the partially purified digestive endonuclease with the chromatin from 6 day-old larvae of the 5th instar resulted in the fragmentation of DNA.The digestive endonuclease was activated by Mg^<2+>/Ca^<2+>, and inhibited by Zn^<2+>, indicating that it will contribute to the process of apoptosis in the midgut during larval-pupal metamorphosis. Less

Report

(3 results)
  • 1993 Annual Research Report   Final Research Report Summary
  • 1992 Annual Research Report
  • Research Products

    (26 results)

All Other

All Publications (26 results)

  • [Publications] Toshiharu FURUSAWA: "Temperature-induced metabolic shifts in diapause and non-diapause eggs of the silkworm,Bombyx mori" Zool.Jb.Physiol.96. 169-180 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Toshiharu FURUSAWA: "Double-stranded ribonuclease activity in the digestive juice and midgut of the silkworm,Bombyx mori" Comparative Biochemistry and Physiology. 104. 795-801 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Toshiharu FURUSAWA: "Stage-dependent limited degradation of vitellin and its localization in the Japanese oak silkworm,Antheraea yamamai,during embryonic development" Comparative Biochemistry and Physiology. 104. 787-794 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Toshiharu FURUSAWA: "Effect of an imidazole compoumd,KK-42,on changes in trehalose content and trehalase activity in the silkglands of the silkworm,Bombyx mori." Applied Entomology and Zoology. 28. 169-176 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Hajime MORI: "Foreign gene expression by a baculovirus with an expanded host range" Journal of General Virology. 73. 1877-1880 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Hajime MORI: "Expression of Bombyx mori CPV polyhedrin in insect cells by using a baculovirus expression vector and its assembry into polyhedra" Journal of General Virology. 74. 99-102 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] FURUSAWA,T: "Temperature-induced metabolic shifts in diapause and non-diapause eggs of the silkworm, Bombyx mori." Zool.Jb.Physiol.96. 169-180 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] FURUSAWA,T: "Stage-dependent limited degradation of vitellin and its localization in the Japanese oaksilkworm, Antheraea yamamai during embryonic development." Comparative Biochemistry and Physiology. 104B. 787-794 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] FURUSAWA,T: "Double-stranded ribonuclease activity in the digestive juice and midgut of the silkworm, Bombyx mori." Comp. Biochem. Physiol.104B. 795-801 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] FURUSAWA,T: "Metabolic biochemistry of yolk protein and amino acid in the eggs of the Japanese oak silkworm, Antherae yamamai, during diapause and embryonic development." In "WILD SILKMOTHS '92" (Akai et al., eds.). (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] FURUSAWA,T: "Effect of an imidazole compound, KK-42, on changes in trehalose content andtrehalase avitivity in the silkglands of the silkworm, Bombyx mori (Lepidoptera : Bombycidae)." Applied Entomology and Zoology. 28. 169-176 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] FURUSAWA,T: "Changes in amino acid pools during embryonic development of the Japanese oak silkworm, Antheraea yamamai (Lipidoptera : Bombycidae)." Appl. Entl Zool.28. 234-237 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] MORI, H.: "Foreign gene expression by a baculovirus with an expanded host range." Journal of General Virology. 73. 1877-1880 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] MORI, H.: "Expression of Bombyx mori CPV polyhedrin in insect cells by using a baculovirus expression vector and its assembly into polyhedra." J.Gen. Virol.74. 99-102 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Toshiharu FURUSAWA: "Temperature-induced metabolic shifts in diapause and non-diapause eggs of the silkworm,Bombyx mori" Zool.Jb.Physiol.96. 169-180 (1992)

    • Related Report
      1993 Annual Research Report
  • [Publications] Toshiharu FURUSAWA: "Double-stranded ribonuclease activity in the digestive juice and midgut of the silkworm,Bombyx mori" Comparative Biochemistry and Physiology. 104. 795-801 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Toshiharu FURUSAWA: "Stage-dependent limited degradation of vitellin and its localization.in the Japanese oak silkworm,Antheraea yamamai,during embryonic development" Comparative Biochemistry and Physiology. 104. 787-794 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Toshiharu FURUSAWA: "Effect of an imidazole compoumd,KK-42 on changes in trehalose content and trehalase activity in the silk-glands of the silkworm.Bombyx mori." Applied Entomology and Zoology. 28. 169-176 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Hajime MORI: "Foreign gene expression by a baculovirus with an expanded host range" Journal of General Virology. 73. 1877-1880 (1992)

    • Related Report
      1993 Annual Research Report
  • [Publications] Hajime MORI: "Expression of Bombyx mori CPV polyhedrin in insect cells,by using a baculovirus expression vector and its assembry into polyhedra" Journal of General Virology. 74. 99-102 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Toshiharu Furusawa: "Double-stranded ribonuclease activity in the digestive juice and midgut of the silkworm,Bumbyx mori." Comp.Biochem.Physiol.(1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] Toshiharu Furusawa: "Stage-dependent limited degradation of vitellin and its localization in Antheraea yamamai during embryonic development." Comp.Biochem.Physiol. (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] Toshiharu Furusawa: "Effect of an imidazule compound,KK-42,on changes in trehalose content and trehalase activity in the silkglands of the silkworm" Appl.Entomol.Zool.28. (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] Toshiharu Furusawa: "Changes in amino acid pools during embryonic development of the Jananese oak silkworm,Antheraea yamamai" Appl.Entomol.Zool. 28. (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] Hajima Mori: "Expression of Bombyx mori cytoplasmic polyhedrosis virus polyhedrin in insect cells by using a baculovirus expression vector,and its assembly into polyhedra" Journal of General Virology. 74. 99-102 (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] Hajima Mori: "Foreign gene expression by a baculovirus vector with an expanded host gene" Journal of General Virology. 73. 1877-1880 (1992)

    • Related Report
      1992 Annual Research Report

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Published: 1992-04-01   Modified: 2016-04-21  

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