オーエスキー病ウイルス前初期蛋白の構造と機能 Mapping of Functional Domains of Aujeszky's Disease Virus Immediate-early Protein

この論文をさがす

著者

    • 田原口, 智士 タハラグチ, サトシ

書誌事項

タイトル

オーエスキー病ウイルス前初期蛋白の構造と機能

タイトル別名

Mapping of Functional Domains of Aujeszky's Disease Virus Immediate-early Protein

著者名

田原口, 智士

著者別名

タハラグチ, サトシ

学位授与大学

北海道大学

取得学位

博士 (獣医学)

学位授与番号

甲第3914号

学位授与年月日

1996-03-25

注記・抄録

博士論文

The 180 kilodalton immediate-early protein (IE180) of Aujeszky's disease virus functions as a strong transactivator of several different promoters and also as a repressor of its own transcription. To map the functional domains of IE180, a series of truncated mutants were prepared. Their transcriptional regulatory activities were analyzed using the chloramphenicol acetyl transferase (CAT) assay for transinduction of a thymidine kinase promoter-CAT gene construct and a glycoprotein X promoter-CAT gene construct and negative regulation of an IE promoter-CAT gene construct. IE180 is localized predominantly in the nuclei of infected cells. To define the nuclear localization signals within IE180, cells transfected with each truncated mutant was analyzed by indirect immunofluorescence. Analysis of mutants truncated from the carboxyterminal end of the 1,460-amino acid polypeptide showed that a polypeptide possessing amino acids 1 to 1,081 retained significant functions of transactivation and autoregulation potential. On the other hand, removing amino acids 1 to 132 resulted in a complete loss of transactivation potential, indicating that the domain responsible for transactivation is located in the amino-terminal end of IE180. Additional amino-terminal truncation up to amino acid 453 did not affect the autoregulation activity, indicating that the region between amino acids 454 and 1081 has autoregulation potential. Indirect immunofluorescence analysis of the truncated mutants showed that two regions including a short sequence of basic amino acid residues (RRKRR) were associated with the nuclear localization of IE180. To assess which region substantially functions as the signal for nuclear localization of IE180 molecule, two deletion mutants lacking each region were constructed. A mutant lacking amino acids 333 to 575 was detected in the nuclei of the transfected cells, whereas the other mutant lacking amino acids 900 to 950 was detected mainly in the cytoplasm. These results suggest that the region of amino acids 900 to 950 is responsible for nuclear localization of IE180.

62p.

Hokkaido University(北海道大学). 博士(獣医学)

目次

  1. 目次 / (0003.jp2)
  2. I.緒言 / p1 (0005.jp2)
  3. II.材料および方法 / p7 (0011.jp2)
  4. 1.細胞およびウイルス / p7 (0011.jp2)
  5. 2.ウイルスの精製 / p7 (0011.jp2)
  6. 3.ウイルスDNAの抽出 / p8 (0012.jp2)
  7. 4.遺伝子クローニングに用いたプラスミド、細菌および培地 / p8 (0012.jp2)
  8. 5.オーエスキー病ウイルス(ADV)遺伝子のクローニング / p9 (0013.jp2)
  9. 6.プラスミドDNAの調製 / p9 (0013.jp2)
  10. 7.プラスミドの構築 / p10 (0014.jp2)
  11. 8.ウエスタンブロット / p13 (0017.jp2)
  12. 9.IEおよび欠失IE遺伝子の無細胞系での転写および翻訳 / p18 (0022.jp2)
  13. 10.クロラムフェニコールアセチルトランスフェラーゼ(CAT)アッセイ / p20 (0024.jp2)
  14. 11.間接蛍光抗体法 / p23 (0027.jp2)
  15. III.結果 / p25 (0029.jp2)
  16. 1.ADV IE遺伝子のサブクローニング / p25 (0029.jp2)
  17. 2.欠失IE遺伝子の作出 / p25 (0029.jp2)
  18. 3.IEおよび欠失IE遺伝子の発現 / p25 (0029.jp2)
  19. 4.IE180および欠失IE180変異体のtkおよびgX遺伝子プロモーターに対する転写活性化作用の解析 / p27 (0031.jp2)
  20. 5.IE180および欠失IE180変異体のADV IE遺伝子プロモーターに対する転写抑制作用の解析 / p31 (0035.jp2)
  21. 6.E180の核への局在に関与する部位の決定 / p34 (0038.jp2)
  22. IV.考察 / p40 (0044.jp2)
  23. V.要約 / p48 (0052.jp2)
  24. 謝辞 / p51 (0055.jp2)
  25. 文献 / p52 (0056.jp2)
  26. 英文抄録 / p61 (0065.jp2)
4アクセス

各種コード

  • NII論文ID(NAID)
    500000132916
  • NII著者ID(NRID)
    • 8000000967500
  • DOI(NDL)
  • 本文言語コード
    • jpn
  • NDL書誌ID
    • 000000297230
  • データ提供元
    • 機関リポジトリ
    • NDL-OPAC
    • NDLデジタルコレクション
ページトップへ