1993 年度実績報告書

エイズ制圧に向けての基礎研究柱1.HIVの分子遺伝学

研究課題

研究課題/領域番号	04269104
研究機関	京都大学
研究代表者	畑中正一京都大学, ウイルス研究所, 教授 (30142300)
研究分担者	塩田達雄東京大学, 医科学研究所, 助手 (00187329) 志田壽利京都大学, ウィルス研究所, 助教授 (00144395) 足立昭夫京都大学, ウィルス研究所, 助教授 (90127043) 品川日出夫大阪大学, 微生物病研究所, 教授 (40029799)
キーワード	HIV-1 / Gag蛋白 / 感染初期 / Env / Rev / eIF5A / V3領域 / 細胞宿主域
研究概要	1.tat,revの多アルギニン領域の機能を探るためにそれらと結合する細胞側因子をaffinity chromatographyにより分離精製し、解析している。pro遺伝子産物の発現と阻害剤及び耐性獲得について分子遺伝学的の研究した(畑中)。2.HIVのコードする各種タンパク質の多量発現、精製、それらの生化学的、免疫学的研究を行った。HIV-1のEnvタンパクのエピトープを解析し、感染者血清と100%反応するエピトープをGag-Envキメラタンパクとして大腸菌で多量産生させて精製する方法を確立した。更に宮崎医大・石川教授らに協力して、組換え体HIVタンパクを抗原として尿中の抗体を検出できる診断法を開発した(品川)。3.HIV-1のgag遺伝子に関するin-frame変異株を20種作製し、その増殖能を調べた。感染サイクルの後期に欠損があるものばかりでなく初期に異常があるものが見出された。しかもこの異常は標的細胞の種類により異なっていた。これらの結果はHIV-1のGag蛋白が感染初期に細胞因子とともに重要な役割を持つことを示している(足立)。4.1)我々が改良した組換えワクシニアによりEnvを生産し、5mg/10^9cellの精製Envを得た。2)HIVの誘導する細胞融合は、CD4^+細胞のマイクロフィラメントが必要であり、又、カゼインキナーゼの役割を示唆した。3)翻訳開始因子eIF5AがRevとRexのコファクターであり、又、標的RNA存在下でRexと結合する因子の存在を示唆した(志田)。5.HIV感染者一名の体内のHIVのenv遺伝子のV3領域を、PCR法で増幅して構造解析を行ったところ、時間の経過とともにV3領域の25番目のアミノ酸にアスパラギン酸からリジンへの変化が生じていた。V3領域を組換えた組換えウイルスを作製したところ、この変化によりHIVの細胞宿主域が拡大するとともに、V3領域の抗原構造も変化することが明かになった(塩田)。

研究成果
(33件)

すべてその他

すべて文献書誌 (33件)

[文献書誌] Nam,S.H.: "Characterization of ribosomal frameshifting for expression of pol gene products of human T-cell leukemia virus type I" J.Virol.67. 196-203 (1993)
[文献書誌] Takahashi,J.B.: "Developmental expression of trk B and low-affinity NGF receptor in the rat retina" Neurosci.Lett.151. 174-177 (1993)
[文献書誌] Takano,E.: "Molecular diversity of calpastatin in human erythroid cells" Arch.Biochem.Biophys.303. 349-354 (1993)
[文献書誌] Adachi,Y.: "Nucleolar targeting signal of Rex protein of human T-cell leukemi virus type I specifically binds to nucleolar shuttle protein B-23" J.Biol.Chem.268. 13930-13934 (1993)
[文献書誌] Nakajima,T.: "Overgrowth of human synoviral cells driven by the human T cell leukemia virus type I tax gene" J.Clin.Invest.92. 186-193 (1993)
[文献書誌] Kubota,S.: "Long cellular repeats flanking a defective HTLV-I provirus:implication for site-targeted integration" Oncogene. 8. 2873-2877 (1993)
[文献書誌] Nosaka,T.: "Cytotoxic activity of rev protein of human immunodeficiency virys type I by nucleolar dysfunction" Exp.Cell Res.209. 89-102 (1993)
[文献書誌] Nosaka,T.: "Novel internal promoter/enhancer of HTLV-I for tax expression" Nucleic Acids Res.21. 5124-5129 (1993)
[文献書誌] Hatanaka,M.: "Nucleolar targeting signal" J.UOEH. 15. 167-197 (1993)
[文献書誌] Kubota,S.: "The origin of human immunodeficiency virus type I rev gene:An evolutionary hypothesis" FEBS Lett.338. 118-121 (1994)
[文献書誌] Ueba,T.: "Expression of fibroblast growth factor receptor-1 in human glioma and meningioma tissues" Neurosurgery. 34. 221-226 (1994)
[文献書誌] El-Farrash,M.A.: "Generation and characterization of a human immunodeficiency virus type 1(HIV-1)mutant resistant to an HIV-1 protease inhibitor" J.Virol.68. 233-239 (1994)
[文献書誌] Morimoto,M.: "Use of the reconbinat chimera proteins,LacZ-Env and Gag-Env,for immunological studies on HIV-1 infection" AIDS Res.Hum.Retroviruses. 9. 971-978 (1993)
[文献書誌] Hashida,S.: "Detection of antibody IgG to HIV-1 in urine by sensitive enzyme immunoassay(immune complex transfer enzyme immunoassay)using recombinant protein as antigens for diagnosis of HIV-1 infection" J.Clin.Lab.Anal.7. 353-364 (1993)
[文献書誌] Hirota,S.: "Sensitive enzyme immunoassay(immune complex transfer enzyme immunoassay)for antibodies to reverse transcriptase(RT)of human immunodeficiency virus type I(HIV-I)using recombinant RT of HIV-I as antigen" Clin.Chem.Enzym.Comms.(in press).
[文献書誌] Hashida,S.: "Anti-HIV-I IgG in urine detected by sensitive enzyme immunoassay(immune complex transfer enzyme immunoassay)using recombinant proteins as antigens for diagnosis of HIV-I infection" Clin.Chem.Enzym.Comms.(in press).
[文献書誌] Miyazawa,T.: "The AP-1 binding site in the feline immunodeficiency virus long terminal repeat is not required for virus replication in feline T lymphocytes" J.Gen.Virol.74. 1537-1580 (1993)
[文献書誌] Sakai,H.: "Integration is essential for efficient gene expression of human immunodeficiency virus type 1" J.Virol.67. 1169-1174 (1993)
[文献書誌] Sakai,H.: "Cell-dependent requirement of human immunodeficiency virus type 1 Vif protein for maturation of virus particles" J.Virol.67. 1663-1666 (1993)
[文献書誌] Sakai,H.: "Compatibility of tat and rev transactivators in the primate lentiviruses" Arch.Virol.129. 1-10 (1993)
[文献書誌] Tomonaga,K.: "The feline immunodeficiency virus ORF-A gene facilitates efficient viral replication in established T-cell lines and peripheral blood lymphocytes" J.Virol.67. 5889-5895 (1993)
[文献書誌] Tomonaga,K.: "Feline immunodeficiency virus gene expression:analysis of the RNA splicing pattern and monocistronic rev mRNA" J.Gen.Virol.74. 2409-2417 (1993)
[文献書誌] Schmelz,M.: "Assembly of vaccinia virus:the second wrapping cisterna is derived from the trans Golgi network" J.Virol.68. 130-147 (1993)
[文献書誌] Kannagi,M.: "Cytotoxic T cell response and expression of the target antigen in HILV-I infection" Leukemia. (in press).
[文献書誌] LeGuern,M.: "Single amino acid change in Tat determine the different rates replication of two sequential HIV-1 isolates" Virology. 195. 441-447 (1993)
[文献書誌] Oka,S.: "Genetic analysis of HIV-1 during rapid progression to AIDS in an apparently healthy man" AIDS Res.Hum.Retroviruses. (in press).
[文献書誌] Hatanaka,M.: "Biology of the Germ Line" Japn Sci.Soc.Press,Karger, 304 (1993)
[文献書誌] Hatanaka,M.: "Molecular Basis of Immune Responses" Academic Press Inc., 287 (1993)
[文献書誌] 畑中正一: "ATLとAIDS" 南江堂, 150 (1994)
[文献書誌] 畑中正一: "ウイルスとの闘い:がん・エイズは克服できるか" NHK出版協会, 128 (1993)
[文献書誌] 川村名子: "エイズ研究の最先端(分担執筆:HIVの制御遺伝子と遺伝子治療)" 羊土社, 166 (1993)
[文献書誌] 川村名子: "HIV感染症・AIDS(分担執筆:HIV遺伝子の構造と機能)" 日本臨牀社, 658 (1993)
[文献書誌] 志田壽利: "エイズ研究の最先端(基礎から臨床へのup-to-dateな分子医学)" 羊土社, 166 (1993)

1993 年度 実績報告書

エイズ制圧に向けての基礎研究 柱1.HIVの分子遺伝学

研究代表者

畑中 正一 京都大学, ウイルス研究所, 教授 (30142300)

研究成果

[文献書誌] Nam,S.H.: "Characterization of ribosomal frameshifting for expression of pol gene products of human T-cell leukemia virus type I" J.Virol.67. 196-203 (1993)

[文献書誌] Takahashi,J.B.: "Developmental expression of trk B and low-affinity NGF receptor in the rat retina" Neurosci.Lett.151. 174-177 (1993)

[文献書誌] Takano,E.: "Molecular diversity of calpastatin in human erythroid cells" Arch.Biochem.Biophys.303. 349-354 (1993)

[文献書誌] Adachi,Y.: "Nucleolar targeting signal of Rex protein of human T-cell leukemi virus type I specifically binds to nucleolar shuttle protein B-23" J.Biol.Chem.268. 13930-13934 (1993)

[文献書誌] Nakajima,T.: "Overgrowth of human synoviral cells driven by the human T cell leukemia virus type I tax gene" J.Clin.Invest.92. 186-193 (1993)

[文献書誌] Kubota,S.: "Long cellular repeats flanking a defective HTLV-I provirus:implication for site-targeted integration" Oncogene. 8. 2873-2877 (1993)

[文献書誌] Nosaka,T.: "Cytotoxic activity of rev protein of human immunodeficiency virys type I by nucleolar dysfunction" Exp.Cell Res.209. 89-102 (1993)

[文献書誌] Nosaka,T.: "Novel internal promoter/enhancer of HTLV-I for tax expression" Nucleic Acids Res.21. 5124-5129 (1993)

[文献書誌] Hatanaka,M.: "Nucleolar targeting signal" J.UOEH. 15. 167-197 (1993)

[文献書誌] Kubota,S.: "The origin of human immunodeficiency virus type I rev gene:An evolutionary hypothesis" FEBS Lett.338. 118-121 (1994)

[文献書誌] Ueba,T.: "Expression of fibroblast growth factor receptor-1 in human glioma and meningioma tissues" Neurosurgery. 34. 221-226 (1994)

[文献書誌] El-Farrash,M.A.: "Generation and characterization of a human immunodeficiency virus type 1(HIV-1)mutant resistant to an HIV-1 protease inhibitor" J.Virol.68. 233-239 (1994)

[文献書誌] Morimoto,M.: "Use of the reconbinat chimera proteins,LacZ-Env and Gag-Env,for immunological studies on HIV-1 infection" AIDS Res.Hum.Retroviruses. 9. 971-978 (1993)

[文献書誌] Hashida,S.: "Detection of antibody IgG to HIV-1 in urine by sensitive enzyme immunoassay(immune complex transfer enzyme immunoassay)using recombinant protein as antigens for diagnosis of HIV-1 infection" J.Clin.Lab.Anal.7. 353-364 (1993)

[文献書誌] Hirota,S.: "Sensitive enzyme immunoassay(immune complex transfer enzyme immunoassay)for antibodies to reverse transcriptase(RT)of human immunodeficiency virus type I(HIV-I)using recombinant RT of HIV-I as antigen" Clin.Chem.Enzym.Comms.(in press).

[文献書誌] Hashida,S.: "Anti-HIV-I IgG in urine detected by sensitive enzyme immunoassay(immune complex transfer enzyme immunoassay)using recombinant proteins as antigens for diagnosis of HIV-I infection" Clin.Chem.Enzym.Comms.(in press).

[文献書誌] Miyazawa,T.: "The AP-1 binding site in the feline immunodeficiency virus long terminal repeat is not required for virus replication in feline T lymphocytes" J.Gen.Virol.74. 1537-1580 (1993)

[文献書誌] Sakai,H.: "Integration is essential for efficient gene expression of human immunodeficiency virus type 1" J.Virol.67. 1169-1174 (1993)

[文献書誌] Sakai,H.: "Cell-dependent requirement of human immunodeficiency virus type 1 Vif protein for maturation of virus particles" J.Virol.67. 1663-1666 (1993)

[文献書誌] Sakai,H.: "Compatibility of tat and rev transactivators in the primate lentiviruses" Arch.Virol.129. 1-10 (1993)

[文献書誌] Tomonaga,K.: "The feline immunodeficiency virus ORF-A gene facilitates efficient viral replication in established T-cell lines and peripheral blood lymphocytes" J.Virol.67. 5889-5895 (1993)

[文献書誌] Tomonaga,K.: "Feline immunodeficiency virus gene expression:analysis of the RNA splicing pattern and monocistronic rev mRNA" J.Gen.Virol.74. 2409-2417 (1993)

[文献書誌] Schmelz,M.: "Assembly of vaccinia virus:the second wrapping cisterna is derived from the trans Golgi network" J.Virol.68. 130-147 (1993)

[文献書誌] Kannagi,M.: "Cytotoxic T cell response and expression of the target antigen in HILV-I infection" Leukemia. (in press).

[文献書誌] LeGuern,M.: "Single amino acid change in Tat determine the different rates replication of two sequential HIV-1 isolates" Virology. 195. 441-447 (1993)

[文献書誌] Oka,S.: "Genetic analysis of HIV-1 during rapid progression to AIDS in an apparently healthy man" AIDS Res.Hum.Retroviruses. (in press).

[文献書誌] Hatanaka,M.: "Biology of the Germ Line" Japn Sci.Soc.Press,Karger, 304 (1993)

[文献書誌] Hatanaka,M.: "Molecular Basis of Immune Responses" Academic Press Inc., 287 (1993)

[文献書誌] 畑中正一: "ATLとAIDS" 南江堂, 150 (1994)

[文献書誌] 畑中正一: "ウイルスとの闘い:がん・エイズは克服できるか" NHK出版協会, 128 (1993)

[文献書誌] 川村名子: "エイズ研究の最先端(分担執筆:HIVの制御遺伝子と遺伝子治療)" 羊土社, 166 (1993)

[文献書誌] 川村名子: "HIV感染症・AIDS(分担執筆:HIV遺伝子の構造と機能)" 日本臨牀社, 658 (1993)

[文献書誌] 志田壽利: "エイズ研究の最先端(基礎から臨床へのup-to-dateな分子医学)" 羊土社, 166 (1993)

1993 年度実績報告書

エイズ制圧に向けての基礎研究柱1.HIVの分子遺伝学

畑中正一京都大学, ウイルス研究所, 教授 (30142300)