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Yeast One Hybrid法によるJC virus複製機構の検討

Research Project

Project/Area Number 08877044
Research Category

Grant-in-Aid for Exploratory Research

Allocation TypeSingle-year Grants
Research Field Experimental pathology
Research InstitutionHokkaido University

Principal Investigator

長嶋 和郎  北海道大学, 医学部, 教授 (50010377)

Co-Investigator(Kenkyū-buntansha) 田中 伸哉  北海道, 医学部, 助手 (70261287)
澤 洋文  北海道, 医学部, 講師 (30292006)
篠原 敏也  北海道大学, 医学部, 助手 (90241306)
Project Period (FY) 1996 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1997: ¥100,000 (Direct Cost: ¥100,000)
Fiscal Year 1996: ¥1,800,000 (Direct Cost: ¥1,800,000)
KeywordsJCウイルス / NF-1 / 転写因子 / Callular tropism / microinjection / regulatory region / PML / 髄液診断 / JC virus / yeast one-hybrid system / X / archetype JC / COS-7 / cellular tropism
Research Abstract

1.Clontech(Palo Alto,CA)社のyeast one-hybrid system)を用いてJC virus NF-1領域に特異的に結合する蛋白を同定した。NF-1配列を用いたスクリーニングで30個の陽性クローンから4個はこれまでにdata baseに登録されていない未知遺伝子であったためにこのDNA配列を確認し、NF-1/Xをcloningし、その発現が脳に多いことを示した。
2.ヒトに無症候性に感染し、常時尿に排泄されている型Archetype JC virusであるCY株(調節領域が原始型)がCOS-7細胞で増殖することを見いだし、この細胞を用いた解析系を樹立した。Mad-1(調節領域がPML型)と調節領域をCYに変えたキメラウイルスCY/Mad-1で感染性を調べ、cellular tropsimに調節領域が関与していることを示した。
3.Microinjection法により非感受性細胞であるCOS7でも細胞膜を越えてvirusを注入するとvirusの増殖が見られることから、virus感染に細胞膜因子が存在することを示唆した。
4.過去2年間に日本国内にて発症が確認されたPMLの5症例のregulatory regionを解析し、4例のPML型と1例のArchetype JCの症例を見い出し、報告した。またPMLの髄液を用いたPCRによる診断方法を確立した。

Report

(2 results)
  • 1997 Annual Research Report
  • 1996 Annual Research Report
  • Research Products

    (14 results)

All Other

All Publications (14 results)

  • [Publications] Shishido Y, et al.: "Assembly of JC virus-like particles in COS7 cells." J Med Virol. 51. 269-277 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kato A, et al.: "Lack of evidence fof the transmission of JC polyoma virus between human populations." Arch Virol. 142. 875-882 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Shinohara T, et al.: "Propagation of the human polyomavirus,JCV,in human neuroblastoma cell lines." Virology. 228. 269-277 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Nagashima K, et al.: "Analysis of and the role of the requlatory region of archetype (CY) and PML type of JC virus in determining cellnlartropism." Neurovirology. 3. 92- (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Nagashima K, et al.: "Analysis of the regulatory region and DNA binding factors of JC virus in determining cellular tropism." Brain Pathol. 7. 1322- (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Sugimoto C, et al.: "Amplification of JC virus regnlatory DNA sequences from cerebrospinal fliud : diagnostic value of PML" Arch Virol. 143. 249-262 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Nukuzuma S, et al.: "Establishment and characterization of a carrier cell culture produting high titers of polyoma JC virus" J Med Virol. 47. 370-377 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Kimura J, et al.: "Inhibitory offects of HIV-1_<8P>120 on myelin formation" Perspectives in Drug Discovery and Design. 5. 17-19 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Shishido Y, et al.: "Assembly of JC virus-like particles in COS 7 cells." J Med Virol. (in press). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Kato A, et al.: "Lack of evidence for the transmission of JC polyomavirus between humnan populations." Arch Virol. (in press). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Obara Y, et al.: "Distribution of herpes simplex virus types 1 and 2 genowies in human spinal ganglia" J Med Virol. (in press). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] 越智 博文,他: "PET にてmethionine集積を認めた進行性多巣性白質脳症の1例" 臨床神経学. 36. 858-863 (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] 長谷川 靖,他: "JCウイルスとPML・実験的脳腫瘍" メジカルビュー社, 14 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 長嶋 和郎: "進行性多巣性白質脳症" 日本臨床社(in press), (1997)

    • Related Report
      1996 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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