プリオン病原体の増殖機構に関する研究

• Project/Area Number: 17780228
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Applied veterinary science
• Research Institution: Osaka University
• Principal Investigator: 作道 章一 Osaka University, 微生物病研究所, 助教 (10397672)
• Project Period (FY): 2005 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥3,100,000 (Direct Cost: ¥3,100,000); Fiscal Year 2007: ¥1,000,000 (Direct Cost: ¥1,000,000); Fiscal Year 2006: ¥1,000,000 (Direct Cost: ¥1,000,000); Fiscal Year 2005: ¥1,100,000 (Direct Cost: ¥1,100,000)
• Keywords: 感染症 / ウイルス / 獣医学

Research Abstract

本研究では、プリオン増殖機構の核である正常型プリオン蛋白質(PrP^C)→異常型プリオン蛋白質(PrP^<Sc>)の変換機構に関与するPrP^C側の要因を解明することを主要な目的とした。これまで、内在性PrP^Cによる阻害効果が障壁となっており、プリオン増殖に必要なPrP^C領域の解析は進展していなかった。研究代表者らはごく最近、プリオン蛋白質(PrP)遺伝子欠損マウスから作製したPrP遺伝子欠損神経細胞株を用い、PrP^Cの機能部位がオクタリピート領域(OR)と疎水性領域(HR)に存在することを示し、さらに、PrP^Cが抗酸化ストレス制御能を持つことを明らかにした。さらに、同様の細胞培養系を用い、一過性PrP^<Sc>増殖に必要なPrP^C領域の同定を行った。その結果、ORが重要であることを示した。平成19年度は、PrP^<Sc>による神経細胞毒性のモデルとして知られるPrPペプチド(PrP106-126)の培地への添加により、PrP再発現PrP遺伝子欠損神経細胞株の血清除去誘導細胞死が促進されることを明らかにした。さらに、PrP再発現細胞やOR欠損PrP発現細胞では、血清除去誘導細胞死がPrP106-126添加により促進されたが、HR欠損PrP発現細胞では促進効果が見られないことが明らかとなった。これらのことから、PrP106-126の神経細胞毒性発揮には細胞側のHRの存在が必要であることが示唆された。

Research Products

• [Journal Article] Normal cytochrome c oxidase activity in prion protein gene-deficient mice (2008)
  • Author(s): Sakudo A, Taniuchi Y, Kobayashi T, Onodera T, Ikuta K
  • Journal Title: Protein Peptide Lett (印刷中)
  • Peer Reviewed
• [Journal Article] GPI-anchor-less human prion protein is secreted and glycosylated but lacks SOD activity (2008)
  • Author(s): Sakudo A, Nakamura I, Tsuji S, Ikuta K
  • Journal Title: Int J Mol Med 21 Pages: 217-222
  • Peer Reviewed
• [Journal Article] PrPSc level and incubation time in a transgenic mouse model expressing Borna disease virus phosphoprotein after intracerebral prion infection (2008)
  • Author(s): Sakudo A, Onodera T, Ikuta K
  • Journal Title: Neurosci Lett 431 Pages: 81-85
  • Peer Reviewed
• [Journal Article] Octapeptide repeat region of prion protein (PrP) is required at an early stage for production of abnormal prion protein in PrP-deficient neuronal cell line (2008)
  • Author(s): Sakudo A, Wu G, Onodera T, Ikuta K
  • Journal Title: Biochem Biophys Res Commun 366 Pages: 657-663
  • Peer Reviewed
• [Journal Article] Reduced response of splenocytes after mitogen-stimulation in the prion protein (PrP) gene-deficient mouse: PrPLP/Doppel production and cerebral degeneration, (2007)
  • Author(s): Kim CK, Hirose Y, Sakudo A, Takeyama N, Kang CB, Taniuchi Y, Matsumoto Y, Itohara S, Sakaguchi S, Onodera T
  • Journal Title: Biochem Biophys Res Commun 358 Pages: 469-474
  • Peer Reviewed
• [Journal Article] Late-onset olfactory deficits and mitral cell loss in mice lacking prion protein with ectopic expression of Doppel (2007)
  • Author(s): Kim CK, Sakudo A, Taniuchi Y, Shigematsu, K, Kang CB, Saeki K, Matsumoto Y, Sakaguchi S, Itohara S, Onodera T
  • Journal Title: Int J Mol Med 20 Pages: 169-176
  • Peer Reviewed
• [Journal Article] Serum withdrawal-induced apoptosis in ZrchI prion protein (PrP) gene-deficient neuronal cell line is suppressed by PrP, independent of Doppel (2007)
  • Author(s): Nishimura T, Sakudo A, Hashiyama Y, Yachi A, Saeki K, Matsumoto Y, Ogawa M, Sakaguchi S, Itohara S, Onodera T
  • Journal Title: Microbiol Immunol 51 Pages: 457-466
  • Peer Reviewed
• [Journal Article] Recent development in prion disease research: diagnostic tools and in vitro cell culture models (2007)
  • Author(s): Sakudo A, Nakamura I, Ikuta K, Onodera T
  • Journal Title: J Vet Med Sci 69 Pages: 329-337
  • NAID: 110006250072
  • Peer Reviewed
• [Journal Article] Prion protein gene-deficient cell lines: Powerful tools for prion biology (2007)
  • Author(s): Sakudo A, Onodera T, Ikuta K
  • Journal Title: Microbiol Immunol 51 Pages: 1-13
  • NAID: 10020281629
  • Peer Reviewed
• [Journal Article] Neurotoxic prion protein (PrP) fragment 106-126 requires the N-terminal half of hydrophobic region of PrP in the PrP-deficient neuronal cell line (2007)
  • Author(s): Sakudo A, Nakamura I, Lee DC, Saeki K, Ikuta K, Onodera T
  • Journal Title: Protein & Peptide Lett 14 Pages: 1-6
  • Peer Reviewed
• [Journal Article] Novel single nucleotide polymorphisms in SP1-binding site of Bovine PRNP promoter in Japanese Black cattle: Impairment of its activity (2007)
  • Author(s): Nakamura I, Xue G, Sakudo A, Saeki K, Matsumoto Y, Ikuta K, and Onodera T
  • Journal Title: Intervirology 50 Pages: 190-196
  • Peer Reviewed
• [Journal Article] Prion protein gene-deficient cell lines : Powerful tools for prion biology. (2007)
  • Author(s): Sakudo A, Onodera T, Ikuta K
  • Journal Title: Microbiol Immunol 51 Pages: 1-13
  • NAID: 10020281629
• [Journal Article] Neurotoxic prion protein (PrP) fragment 106-126 requires the N-terminal half of hydrophobic region of PrP in the PrP-deficient neuronal cell line (2007)
  • Author(s): Sakudo A, Nakamura I, Lee DC, Saeki K, Ikuta K, Onodera T
  • Journal Title: Protein ＆ Peptide Lett 14 Pages: 1-6
• [Journal Article] Novel single nucleotide polymorphisms in SP1-binding site of Bovine PRNP promoter in Japanese Black cattle : Impairment of its activity. (2007)
  • Author(s): Nakamura I, Xue G, Sakudo A, Saeki K, Matsumoto Y, Ikuta K, Onodera T
  • Journal Title: Intervirology 50 Pages: 190-196
• [Journal Article] Bovine Spongiform Encephalopathy in Japan : History and Recent Studies on Oxidative Stress in Prion Diseases. (2006)
  • Author(s): Onodera T, Sakudo A, Wu G, Saeki K
  • Journal Title: Microbiol Immunol 50 Pages: 565-578
  • NAID: 10018168090
• [Journal Article] Ex vivo tissue discrimination by visible and near-infrared spectra with chemometrics (2006)
  • Author(s): Sakudo A, Tsenkova R, Tei K, Morita H, Ikuta K, Onodera T
  • Journal Title: J Vet Med Sci 68 Pages: 1375-1378
  • NAID: 130000447991
• [Journal Article] Comparison of Vibration Mode of Metals in HN03 by Partial Least Squares Regression Analysis of Near-Infrared Spectra. (2006)
  • Author(s): Sakudo A, Tsenkova R, Tei K, Onozuka T, IKUTA K, Yoshimura E, Onodera T
  • Journal Title: Biosci Biotechnol Biochem 70 Pages: 1578-1583
• [Journal Article] Octapeptide repeat region and N-terminal half of hydrophobic region of prion protein(PrP) mediate PrP-dependent activation of superoxide dismutase. (2006)
  • Author(s): Sakudo A, Lee DC, Nishimura T, Li S, Tsuji S, Nakamura T, Matsumoto Y, Saeki K, Itohara S, Ikuta K, Onodera T
  • Journal Title: Biochem Biophys Res Commun 326・3 Pages: 600-606
• [Journal Article] Near-infrared spectroscopy : Promising diagnostic tool for viral infections. (2006)
  • Author(s): Sakudo A, Suganuma Y, Kobayashi T, Onodera T, Ikuta K
  • Journal Title: Biochem Biophys Res Commun 341・2 Pages: 279-284
• [Journal Article] BSEの分子生物学(Molecular biology of bovine spongiform encephalopathy) (2006)
  • Author(s): 小野寺節, 作道章一, 佐伯圭一, 谷内洋次郎, 中村出
  • Journal Title: 化学療法の領域(医療ジャーナル) 22・1 Pages: 49-54
• [Journal Article] Recent advances in clarifying prion protein functions using knockout mice and derived cell lines. (2006)
  • Author(s): Sakudo A, Onodera T, Suganuma Y, Kobayashi T, Saeki K, Ikuta k
  • Journal Title: Mini-Rev Med Chem (印刷中)
• [Journal Article] Fusion of Doppel to octapeptide repeat and N-terminal half of hydrophobic region of prion protein confers resistance to serum deprivation. (2006)
  • Author(s): Lee DC, Sakudo A, Kim CK, Nishimura T, Saeki K, Matsumoto Y, Yokoyama T, Chen SG, Itohara S, Onodera T
  • Journal Title: Microbiol Immunol (印刷中)
  • NAID: 10017327601
• [Journal Article] Prp cooperates with STLL to regulate SOD activity in PrP-deficient neuronal cell line. (2005)
  • Author(s): Sakudo A, Lee DC, Li S, Nakamura I, Matsumoto Y, Saeki K, Itohara S, Ikuta K, Onodera T
  • Journal Title: Biochem Biophys Res Commun 328・1 Pages: 14-19
• [Journal Article] Infection route-independent accumulation of splenic abnormal prion protein. (2005)
  • Author(s): Inoue Y, Yamakawa Y, Sakudo A, Kinumi T, Nakamura Y, Matsumoto Y, Saeki K, Kamiyama T, Onodera T, Nishijima M
  • Journal Title: Jpn J Infect DiS 58・2 Pages: 78-82
  • NAID: 40006702870
• [Journal Article] Cell-autonomous PrP-Doppel interaction regulates apoptosis in PrP gene-deficient neuronal cells. (2005)
  • Author(s): Sakudo A, Lee DC, Nakamura.l, Taniuchi Y, Saeki K, Matsumoto Y, Itohara S, Ikuta K, Onodera T
  • Journal Title: Biochem Biophys Res Commun 333・2 Pages: 448-454
• [Journal Article] A novel diagnostic method for human immunodeficiency virus type-1 in plasma by near-infrared spectroscopy. (2005)
  • Author(s): Sakudo A, Tsenkova R, Onozuka T, Morito K, Li S, Warachit J, Iwabu Y, Li G, Onodera T, Ikuta K
  • Journal Title: Microbiol Immunol 49・7 Pages: 695-701
  • NAID: 10016568061
• [Journal Article] 正常型プリオンタンパクの抗アポトーシス機能における構造的な機能中心の解析 (2005)
  • Author(s): 李得燦, 作道章一, 佐伯圭一, 小野寺節
  • Journal Title: 獣医生化学(Veterinary biochemistry) 42 Pages: 19-23
  • NAID: 10024393341
• [Presentation] RK13細胞発現系を用いた組換え分泌性ヒトプリオンタンパク質の発現および性状解析 (2007)
  • Author(s): 作道章一
  • Organizer: 第55回日本ウイルス学会学術集会
  • Place of Presentation: 札幌
• [Remarks]
  • URL: http://virology.biken.osaka-u.ac.jp/
• [Remarks]
  • URL: http://www.biken.osaka-u.ac.jp/act/act_ikuta.php