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STUDIES ON THE STRUCTURE AND FUNCTION OF NONSTRUCTURAL PROTEINS OF ROTAVIRUS

Research Project

Project/Area Number 07457080
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Virology
Research InstitutionSAPPORO MEDICAL UNIVERSITY

Principal Investigator

TANIGUCHI Koki  Sapporo Med.Univ., Dept.of Hygiene, Associate Professor, 医学部, 助教授 (40094213)

Co-Investigator(Kenkyū-buntansha) KOJIMA Kazunobu  Sapporo Med.Univ., Dept.of Hygiene, Instructor, 医学部, 助手 (20264517)
KOBAYASHI Nobumichi  Sapporo Med.Univ., Dept.of Hygiene, Associate Professor, 医学部, 助教授 (80186759)
URASAWA Tomoko  Sapporo Med.Univ., Dept.of Hygiene, Professor, 保健医療学部, 教授 (90045378)
Project Period (FY) 1995 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥7,700,000 (Direct Cost: ¥7,700,000)
Fiscal Year 1997: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1996: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1995: ¥3,900,000 (Direct Cost: ¥3,900,000)
KeywordsROTAVIRUS / BACULOVIRUS / VACCINE / DIARRHEA / ARTIFICIAL PARTICLE / SEQUENCE / DELETION / NONSENSE MUTATION / ヒトロタウイルス / バキュロウイルス発現系 / 下痢発症 / リアソ-タント / プラークサイズ / 非構造蛋白 / NSP1 / 非構造蛋白質 / リアレンジメント / 亜鉛フィンガーモチーフ
Research Abstract

1.We isolaed subclones producing large, medium, or small plaques from each of A5-13 expressing normal NSP1, A5-10 encoding omly 40 amino acid-NSP1, and A5-16 having NSP1 gene with 500 bp deletion. All of the clones replicated well in MA-104 cells, and induced dirrhea in suckling mice by oral administration. These results indicate that the insertion of foreign gene into NSP1 gene will be possible when the system of artificial infectious virus particles is established.
2.We first determined the complete nucleotide sequence of the genome of human ratavirus (strain KU).
3.All of the 6 structural proteins VP1-VP4, VP6, VP7 and 4 of 5 non-structural proteins of human rotavirus were expressed in baculovirus expression system. The produre for purification of his-tagged nonstructural proteins was established, and this enabled the precise examination of the function of nonstructural proteins.
4.Self-assembled, artificial and empty single-shelled particles or double-shelled particules were obtained by coexpressions of VP2 and VP6, or VP2, VP6 and VP7. In addition, the coexpression of VP1, VP3, VP6, VP7, and VP4 as well as VP2 resulted in the formation of artificial virus-like particles lacking genome. These artificial empty particles have the value as a value as a vaccine candidate and a tool for studying the immunity of rotavirus infection.
5.In the infection experiments by oral inoculation to suckling mice, we observed the diarrhea in the mice administered with G1, G3, G4, G8, and G12 human rotavirus strains. This finding show that it is possible to carry out the rotavirus infection-protection experiments in mice by using human rotaviruses.

Report

(4 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • 1995 Annual Research Report
  • Research Products

    (32 results)

All Other

All Publications (32 results)

  • [Publications] K.Taniguchi: "Nondefective rotavirus mutants with an NSP1 gene which has a deletion of 500 nucletides,including a cysteine-rich zinc finger motif-encoding region(nucleotides 156 to 248),or which has a nonsense codon at nucleotides 153 to 155" J.Virol.70. 4125-4130 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] K.Kojima: "Species-specific and interspecies relatedness of NSP1 sequences in human,porcine,feline,and equine rotavirus strains" Arch.Virol.141. 1-12 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Y.Pongsuwanna: "Serological and genomic characterization of porcine rotaviruses in Thailand:detection of a G10 porcine rotavirus" J.Clin.Microbiol.34. 1050-1057 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] K.Kojima: "Sequence analysis of normal and rearranged NSP5 genes from human rotavirus strains isolated in nature:implications for the occurrence of the rearrangement at the step of plus strand synthesis." Virology. 224. 446-452 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] K.Taniguchi: "Structure and function of rotavirus NSP1" Arch.Virol.12. 53-58 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] H.Wu: "Serological and genomic characterizations of human rotaviruses detected in China" J.Med.Virol.(in press). (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] K.Taniguchi, K.Kojima, S.Urasawa: "Nondefective rotavirus mutants with an NSPI gene which has a deletion of 500 nucleotides, including a cysteine-rich zinc finger motif-encoding region (nucleotides 156 to 248), or which has a nonsense codon at nucleotides 153 to 155." J.Virol. 70. 4125-4130 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] A.Fukushima.Y.C.Yoo, K.Yoshimatsu, K.Matsuzawa, M.Tamura, S.Tono-oka, K.Taniguchi, S.Urasawa, J.Arikawa, I.Azuma: "Effect of MDP-Lys (L18) as a mucosal immunoadjuvant on protection of mucosal infections by Sendai virus and rotavirus." Vaccine. 14. 485-491 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] K.Kojima, K.Taniguchi, N.Kobayashi: "Species-specific and interspecies relatedness of NSPI sequences in human, porcine, feline, and equine rotavirus strains." Arch.Virol.141. 1-12 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] N.Kobayashi, J.Okada, K.Taniguchi, T.Urasawa, S.Urasawa: "Selection of rotavirus VP7 gene in the genetic background of simian rotavirus SA11 : implications for rotavirus reassortant vaccine development." Antiviral Res.31. 185-190 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] N.Kobayashi, K.Taniguchi, K.KOjima, T.Urasawa, S.Urasawa: "G(VP7)serotype-dependent preferential VP7 gene selection detected in the genetic background of simian rotavirus SA11" Arch.Virol.141. 1167-1176 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Y.Pongsuwanna, K.Taniguchi, M.Chiwakul, T.Urasawa, F.Wakasugi, C.Jayavasu, S.Urasawa: "Serological and genomic characterization of porcine rotaviruses in Thailand : detection of a G10 porcine rotavirus." J.Clin.Microbiol.34. 1050-1057 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] K.Kojima, K.Taniguchi, T.Urasawa, S.Urasawa: "Sequence analysis of normal and rearranged NSP5 genes from human rotavirus strains isolated in nature : implications for the occurrence of the rearrangement at the step of plus strand synthesis." Virology. 224. 446-452 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] K.Taniguchi, K.Kojima, N.Kobayashi, T.Urasawa, S.Urasawa: "Structure and function of rotavirus NSP1." Arch.Virol.12[Supple]. 53-58 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] H.Wu, K.Taniguchi, T.Urasawa, S.Urasawa: "Serological and genomic characterization of human rotaviruses detected in China" J.Med.Virol.(in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] J.Okada.N.KObayashi, K.Taniguchi, S.Urasawa: "Preferential selection of heterologous G3-VP7 gene in the genetic background of simian rotavirus SA11 detected by using a homotypic single-VP7 gene substitution reassortant." Autiviral Res.(in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Wu Huixia: "Serological and genomic characterizations of human rotaviruses detected in China." J.Med.Virol.(in press). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Okada Junichi: "Preferential selection of heterologous G3-VP7 gene in the genetic'background of simian rotavirus SA11 detected by using a homotypic single-VP7 gene-substitution reassortant" Antiviral Res.(in press). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Sakawaki Hiromi: "Occurrence of a single type strain(G3,P[12])of equine rotavirus in foals on a racehorse farm from 1986 to 1993." Res.Bull.Obihiro Univ.20. 103-111 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 谷口 孝喜: "下痢症ウイルス研究の進展" 実験医学. 15. 211-218 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Koki Taniguchi: "Non-defective rotavirus mutants with NSP1 gene which has a deletion of 500 nucleotides including a cysteine-rich Zn finger motif encoding (nucleotide no.156-248) or which has a nonsense codon at nucleotide no.153-155." J.Virol.70. 4125-4130 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Kojima Kazunobu: "Sequence analysis of normal and rearranged NSP5 genes from human rotavirus strains isolated in nature : implication for the occurrence of the rearrangement at the step of plus strand synthesis." virology. 224. 446-452 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] kojima Kazunobu: "Species-specific and interspecies relatedness of NSP1 sequences in human,porcine,bovine,eline,and equine rotavirus strains." Arch.Virol.141. 1-12 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Kobayashi Nobumichi: "G (VP7) serotype-dependent preferential VP7 gene selection detected in genetic background of simian rotavirus SA11" Arch.Virol.141. 1167-1176 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Kobayashi Nobumichi: "Slection of rotavirus VP7 gene in the genetic background of simian rotavirus SA11 : implication for rotavirus reassortant vaccine development." Antiviral Res.31. 185-190 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Pongsuwanna Yaowapa: "Serological and genomic characterization of porcine rotaviruses in Thailand : detection of a G10 porcine rotavirus." J.Clin.Microbiol.34. 1050-1057 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Koki Taniguchi: "Non-defective rotavirus mutants with NSP1 gene which has a deletion of 500 nucleotides including a cysteine-rich Zn finger motif encoding region (nucleotide no.156-248)or which has a nonsense codon at nucleotide no.153-155." J.Virol.(in press). (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] Koki Taniguchi: "Diversity in rotavirus genomes." Sem.Virol.6. 123-131 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Kazunobu Kojima: "Species-specific and interspecies relatedness of NSP1 sequences in human,porcine,bovine,feline,and equine rotavirus strains." Arch.Virol.141(in press). (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] Nobumichi Kobayashi: "Non-random selection of gene segment 3 and random selection of gene segment 5 observed in reassortants generated in vitro between rotavirus SA11 and RRV." Res.Virol.146. 53-59 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Nobumichi Kobayashi: "Preferential selection of specific rotavirus gene segments in coinfection and multiple passages with reassortant viruses and their parental strain." Res.Virol.146. 333-342 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Yaowapa Pongsuwanna: "Serological and genomic characterization of porcine rotaviruses in Thailand:Detection of a G10 porcine rotavirus" J.Clin.Microbiol.(in press). (1996)

    • Related Report
      1995 Annual Research Report

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Published: 1995-04-01   Modified: 2016-04-21  

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