Comprehensive understanding of the virulence mechanisms of Entamoeba histolytica

• Project/Area Number: 17390124
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Parasitology (including Sanitary zoology)
• Research Institution: Gunma University
• Principal Investigator: NOZAKI Tomoyoshi GUNMA UNIVERSITY, Department of Parasitology, Professor, 大学院・医学系研究科, 教授 (60198588)
• Co-Investigator(Kenkyū-buntansha): SAITO Yumiko (NAKANO Yumiko) National Institute of Infectious Diseases, Department of Parasitology, Senior Research Scientist, 寄生動物部, 主任研究官 (30321764)
• Project Period (FY): 2005 – 2006
• Project Status: Completed (Fiscal Year 2006)
• Budget Amount: ¥14,600,000 (Direct Cost: ¥14,600,000); Fiscal Year 2006: ¥5,600,000 (Direct Cost: ¥5,600,000); Fiscal Year 2005: ¥9,000,000 (Direct Cost: ¥9,000,000)
• Keywords: Entamoeba histolytica / Pathogenesis / Cysteine protease / Transcriptome / Vesicular trafficking / Infectious diseases / Parasite / 腸管原虫 / 病原性 / 網羅的解析 / 寄生

Research Abstract

For comprehensive understanding of pathogenesis of Entamoeba histolytica, transcriptome analysis using DNA microarrays was conducted. The array covers 7712 genes out of 9435 genes in the TIGR genome database. Approximately 80% of genes were expressed in the reference strain axenically cultivated. We also established highly-virulent or attenuated lines by passaging the hamster liver or continuously cultivating in the axenic medium. These lines showed remarkable differences in both in vitro and in vivo pathogenicity. Transcriptome analysis of these lines revealed approximately 200 genes were differentially expressed, about 30 or 170 of which were up- or down-regulated in the highly-virulent line. We also discovered a new family of intrinsic inhibitor, named ICP, which regulated transport and activity of the major virulence factor cysteine proteases (CP). E. histolytica possesses ICP1 and ICP2, located in the cytosol and lysosomes/phagosomes, respectively. Both ICP1 and ICP2 bound to the major CPs (CP1,2,5)) in vivo, and inhibited these CPs in vitro. CP existed 1000-fold in excess to ICP. The amoeba transformants overexpressing ICP1 or ICP2 showed 60-90% reduction of both intracellular and secreted CP activity. These data indicate that ICPs bind to and regulate CP by binding to CPs in distinct compartments and further leading to degradation.

Research Products

• [Journal Article] Kinetics and strain variation of phagosome proteins of Entamoeba histolytica by proteomic analysis. (2006)
  • Author(s): Okada, M., Huston, C.D., Oue, M., Mann, B.J., Petri, Jr., W.A., Kita, K., Nozaki, T.
  • Journal Title: Mol. Biochem. Parasitol. 145 Pages: 171-183
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Entamoeba histolytica : Differences in phagosome acidification and degradation between attenuated and virulent strains. (2006)
  • Author(s): Mitra, B.N., Kobayashi, S., Saito-Nakano, Y, Nozaki, T.
  • Journal Title: Exp. Parasitol. 114 Pages: 57-61
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Two cysteine protease inhibitors, EhICP1 and 2, localized in distinct compartments, negatively regulate secretion in Entamoeba histolytica. (2006)
  • Author(s): Sato, D., Nakada-Tsukui, K., Okada, M., Nozaki, T.
  • Journal Title: FEBS Lett. 580 Pages: 5306-5312
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Impact of intestinal colonization and invasion on the Entamoeba histolytica transcriptome. (2006)
  • Author(s): Gilchrist, C.A., Houpt, E., Trapaidze, N., Fei, Z., Crasta, O., Asgharpour, A., Evans, C., Martino-Catt, S., Baba, D.J., Stroup, S., Hamano, S., Ehrenkaufer, G., Okada, M., Singh, U., Nozaki T., Mann, B.J., Petri, Jr., W.
  • Journal Title: Mol. Biochem. Parasitol. 147 Pages: 163-176
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] 寄生虫への進化 : 驚くべき寄生虫のメンブレントラフィック (2006)
  • Author(s): 野崎智義, 中野由美子
  • Journal Title: 細胞工学 25 Pages: 672-676
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Kinetics and strain variation of phagosome proteins of Entamoeba histolytica by proteomic analysis. (2006)
  • Author(s): Okada, M., Huston, C.D., Oue, M., Mann, B.J., Petri, Jr., W.A., Kita, K., Nozaki, T.
  • Journal Title: Mol.Biochem.Parasitol. 145 Pages: 171-183
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Entamoeba histolytica : Differences in phagosome acidification and degradation between attenuated and virulent strains. (2006)
  • Author(s): Mitra, B.N., Kobayashi, S., Saito-Nakano, Y, Nozaki, T.
  • Journal Title: Exp.Parasitol. 114 Pages: 57-61
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Impact of intestinal colonization and invasion on the Entamoeba histolytica transcriptome. (2006)
  • Author(s): Gilchrist, C.A., Houpt, E., Trapaidze, N., Fei, Z., Crasta, O., Asgharpour, A., Evans, C., Martino-Catt, S., Baba, D.J., Stroup, S., Hamano, S., Ehrenkaufer, G., Okada, M., Singh, U., Nozaki, T., Mann, B.J., Petri, Jr., W.
  • Journal Title: Mol.Biochem.Parasitol. 147 Pages: 163-176
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Two cysteine protease inhibitors, EhlCP1 and 2, localized in distinct compartments, negatively regulate secretion in Entamoeba histolytica. (2006)
  • Author(s): Sato, D., Nakada-Tsukui, K., Okada, M., Nozaki,T.
  • Journal Title: FEBS Lett. 580 Pages: 5306-5312
• [Journal Article] Impact of intestinal colonization and invasion on the Entamoeba histolytica transcriptome. (2006)
  • Author(s): Gilchrist, C.A., Houpt, E., Trapaidze, N., Fei, Z., Crasta, O., Asgharpour, A., Evans, C., Martino-Catt, A., Baba, D.J., Stroup, S., Hamano, S., Ehrenkaufer, G., Okada, M., Singh, U., Nozaki, T., Mann, B.J., Petri,Jr., W.
  • Journal Title: Mol. Biochem. Parasitol. 147 Pages: 163-176
• [Journal Article] Entamoeba histolytica : Differences in phagosome acidification and degradation between attenuated and virulent strains. (2006)
  • Author(s): Mitra, B.N., Kobayashi, S., Saito-Nakano, Y, Nozaki, T.
  • Journal Title: Exp.Parasitol. (In press)
• [Journal Article] Identification and gene expression analysis of a large family of transmembrane kinases related to the Gal/GalNAc lectin in Entamoeba histolytica. (2005)
  • Author(s): Beck, D.L., Boettner, D., Dragulev, B., Ready, L., Mackey, A.J., Nozaki, T., Pearson, W.R., Petri, Jr., W.A.
  • Journal Title: Eukaryot. Cell 4 Pages: 722-732
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Proteomic analysis of phagocytosis in the enteric protozoan parasite Entamoeba histolytica. (2005)
  • Author(s): Okada, M., Huston, C.D., Mann, B.J., Petri, Jr., W.A., Kita, K., Nozaki, T.
  • Journal Title: Eukaryot. Cell 4 Pages: 827-831
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] The diversity of Rab small GTPases in Entamoeba histolytica. (2005)
  • Author(s): Saito-Nakano, Y., Loftus, B.J., Hall, N., Nozaki, T.
  • Journal Title: Exp. Parasitol. 110 Pages: 244-252
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Differences in morphology of phagosomes and kinetics of acidification and degradation in phagosomes between the pathogenic Entamoeba histolytica and the non-pathogenic Entamoeba dispar. (2005)
  • Author(s): Mitra, B.N., Yasuda, T., Kobayashi, S., Satio-Nakano, Y., Nozaki, T.
  • Journal Title: Cell Motil. Cytoskeleton 62 Pages: 84-99
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] A retromerlike complex is a novel Rab7 effector that is involved in the transport of the virulence factor cysteine protease in the enteric protozoan parasite Entamoeba histolytica. (2005)
  • Author(s): Nakada-Tsukui, K., Saito-Nakano, Y., Ali, V., Nozaki, T.
  • Journal Title: Mol. Biol. Cell 16 Pages: 5294-5303
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Identification and gene expression analysis of a large family of transmembrane kinases related to the Gal/GaINAc lectin in Entamoeba histolytica (2005)
  • Author(s): Beck, D.L., Boettner, D., Dragulev, B., Ready, L., Mackey, A.J., Nozaki, T., Pearson, W.R., Petri, Jr., W.A.
  • Journal Title: Eukaryot.Cell 4 Pages: 722-732
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Proteomic analysis of phagocytosis in the enteric protozoan parasite Entamoeba histolytica. (2005)
  • Author(s): Okada, M., Huston, C.D., Mann, B.J., Petri, Jr., W.A., Kita, K., Nozaki, T.
  • Journal Title: Eukaryot.Cell 4 Pages: 827-831
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] The diversity of Rab small GTPases in Entamoeba histolytica. (2005)
  • Author(s): Saito-Nakano, Y., Loftus, B.J., Hall, N., Nozaki, T.
  • Journal Title: Exp.Parasitol. 110 Pages: 244-252
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Differences in morphology of phagosomes and kinetics of acidification and degradation in phagosomes between the pathogenic Entamoeba histolytica and the non-pathogenic Entamoeba dispar. (2005)
  • Author(s): Mitra, B.N., Yasuda, T., Kobayashi, S., Satio-Nakano, Y., Nozaki, T.
  • Journal Title: Cell Motil.Cytoskeleton 62 Pages: 84-99
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] A retromerlike complex is a novel Rab7 effector that is involved in the transport of the virulence factor cysteine protease in the enteric protozoan parasite Entamoeba histolytica. (2005)
  • Author(s): Nakada-Tsukui, K., Saito-Nakano, Y., Ali, V., Nozaki, T.
  • Journal Title: Mol.Biol.Cell 16 Pages: 5294-5303
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Identification and gene expression analysis of a large family of transmembrane kinases related to the Gal/GalNAc lectin in Entamoeba histolytica. (2005)
  • Author(s): Beck, D.L., Boettner, D., Dragulev, B., Ready, L., Mackey, A.J., Nozaki, T., Pearson, W.R., Petri, Jr., W.A.
  • Journal Title: Eukaryot.Cell 4 Pages: 722-732