Roles of mitochondria in iron metabolism

• Project/Area Number: 16370061
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Functional biochemistry
• Research Institution: Kyoto University
• Principal Investigator: IWAI Yuko Kyoto University, Graduate School of Biostidies, Associate professor, 生命科学研究科, 助教授 (10281726)
• Co-Investigator(Kenkyū-buntansha): IWAI Kazuhiro Osaka City University, Graduate School of Medicine, Professor, 大学院医学研究科, 教授 (60252459)
• Project Period (FY): 2004 – 2006
• Project Status: Completed (Fiscal Year 2006)
• Budget Amount: ¥15,400,000 (Direct Cost: ¥15,400,000); Fiscal Year 2006: ¥4,100,000 (Direct Cost: ¥4,100,000); Fiscal Year 2005: ¥4,900,000 (Direct Cost: ¥4,900,000); Fiscal Year 2004: ¥6,400,000 (Direct Cost: ¥6,400,000)
• Keywords: mitochondria / iron metabolism / iron-sulfur cluster / knock-out / Saccharomyces cerevisiae / 鉄-硫黄ラスター / DT40 / 鉄 / 鉄ー硫黄クラスター / トリBリンパ細胞

Research Abstract

Mitochondria have been shown to be the organelle to play a central role in iron metabolism by generating prosthetic groups of iron such as heme or iron-sulfur (Fe-S) cluster. However, transport of iron into mitochondria or export of iron prosthetic groups from the organelle as well as the roles of mitochondria play in the control of cellular iron metabolism have not been clarified yet. Here we analyzed roles of mitochondria play in iron metabolism.
Aftlp is an iron-responsive transcriptional activator that plays a central role in maintaining iron homeostasis in Saccharomyces cerevisiae and its activity is regulated primarily by iron-induced nuclear export of the protein. We have shown that Msn5p is the nuclear export receptor (exportin) for Aftlp. Msn5p recognizes Aftlp in the iron-replete condition and the iron-induced intermolecular interaction of Aftlp are essential for its recognition by Msn5p. Mutation of Cys291 of Aftlp to Phe, which causes Aftlp to be retained in the nucleus and results in constitutive activation of Aftl-target genes, disrupts the intermolecular interaction of Aftlp.
We also analyzed the roles of the molecules involved in Fe-S cluster biogenesis and transport using chicken DT40 B cells. Disruption of the molecules involved in Fe-S biogenesis in mitochondria abolishes Fe-S loading to IRP1 and renders the protein to the IRE binding form. However, disruption of the molecules involved in cytoplasmic Fe-S assembly leads IRP1 to degradation by the proteasome in condition of iron-replete although Fe-S cluster cannot bind to the protein in those cells. These results suggest that the IRE-binidng activity of IRP1 is regulated via Fe-S dependent and independent pathways.

Research Products

• [Journal Article] Mechanism Underlying the Iron-Dependent Nuclear Export of the Iron-Responsive Transcription Factor Aft1p in Saccharomyces Cervisiae (2007)
  • Author(s): Fujiwara N., Ueta R., Iwai K., Yamaguchi-Iwai Y.
  • Journal Title: Molecular Biology of the Cell (In revision)
• [Journal Article] Control of mitochondrial outer membrane permeabilization and Bcl-xL levels by thioredoxin 2 in DT40 cells. (2006)
  • Author(s): Wang, D., Masutani, H., Oka, S.I., Tanaka, T., Yamaguchi-Iwai, Y., Nakamura.H., Yodoi, J.
  • Journal Title: J. Biol. Chem. 281 Pages: 7384-7391
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Zinc transport complexes contribute to the homeostatic maintenance of secretory pathway function in vertebrate cells. (2006)
  • Author(s): Ishihara, K, Yamazaki, T., Ishida, Y., Suzuki, T., Oda, K., Nagao, M., Yamaguchi-Iwai, Y., Kambe, T.
  • Journal Title: J. Biol. Chem 281 Pages: 17743-17750
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Sequence similarity and functional relationship among eukaryotic ZIP and CDF transporters. (2006)
  • Author(s): Kambe, T., Suzuki, T., Nagao, M., Yamaguchi-Iwai, Y.
  • Journal Title: Genomics Proteomics Bioinformatics. 4 Pages: 1-9
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Control of mitochondrial outer membrane permeabilization and Bcl-xL levels by thioredoxin 2 in DT40 cells. (2006)
  • Author(s): Wang, D., Masutani, H., Oka, S.I., Tanaka, T., Yamaguchi-Iwai, Y., Nakamura, H., Yodoi, J.
  • Journal Title: J.Biol.Chem. 281 Pages: 7384-7391
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Zinc transport complexes contribute to the homeostatic maintenance of secretory pathway function in vertebrate cells (2006)
  • Author(s): Ishihara, K, Yamazaki, T., Ishida, Y., Suzuki, T., Oda, K., Nagao, M., Yamaguchi-Iwai, Y., Kambe, T.
  • Journal Title: J. Biol. Chem. 281 Pages: 17743-17750
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Sequence similarity and functional relationship among eukaryotic ZIP and CDF transporters. (2006)
  • Author(s): Kambe, T., Suzuki, T., Nagao, M., Yamaguchi- Iwai, Y.
  • Journal Title: Genomics Proteomics Bioinformatics. 4 Pages: 1-9
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Zinc Transport Complexes Contribute to the Homeostatic Maintenance of Secretory Pathway Function in Vertebrate Cells (2006)
  • Author(s): Ishihara K., Yamazaki T., Ishida Y., Suzuki T., Oda K., Nagao, M., Yamaguchi-Iwai Y., Kambe T.
  • Journal Title: The Journal of Biological Chemistry 281 Pages: 17743-17750
• [Journal Article] Sequence similarity and functional relationship among eukaryotic ZIP and CDF transporters (2006)
  • Author(s): Kambe T., Suzuki T., Nagao M., Yamaguchi-Iwai Y.
  • Journal Title: Genomics, Proteomics ＆ Bioinformatics 4 Pages: 1-9
• [Journal Article] Control of mitochondrial outer membrane permeabilization and Bcl-xL levels by thioredoxin 2 in DT40 cells (2006)
  • Author(s): Wang, D., Masutani, H., Oka, S.I., Tanaka T., Yamaguchi-Iwai, Y., Nakamura, H., Yodoi, J.
  • Journal Title: The Journal of Biological Chemistry 281 Pages: 7384-7391
• [Journal Article] Control of mitochondrial outer membrane permeabilization and Bcl-xL levels by thioredoxin 2 in DT40 cells. (2006)
  • Author(s): Wang, D., Masutani, H., Oka, S.I., Tanaka, T., Yamaguchi-Iwai, Y., Nakamura, H., Yodoi, J.
  • Journal Title: J.Biol.Chem. 281(in press)
• [Journal Article] Systematic yeast synthetic lethal and synthetic dosage lethal screens identify genes required for chromosome segregation. (2005)
  • Author(s): Measday, V., Baetz, K., Guzzo, J., Yuen, K., Kwok, T., Sheikh, B., Ding, H., Ueta, R., Hoac, T., Cheng, B., Pot, I., Tong, A., Yamaguchi-Iwai, Y., Boone, C., Hieter, P., Andrews.B.
  • Journal Title: Proc. Natl. Acad. Sci. USA 102 Pages: 13956-13961
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Zinc transporters, ZnT5 and ZnT7, are required for the activation of alkaline phosphatases, zinc-requiring enzymes that are glycosylphosphatidylinositol-anchored to the cytoplasmic membrane. (2005)
  • Author(s): Suzuki, T., Ishihara, K., Migaki, H., Matsuura, W., Kohda, A., Okumura, K., Nagao, M., Yamaguchi-Iwai, Y., Kambe, T.
  • Journal Title: J Biol Chem 280 Pages: 637-643
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Two different zinc transport complexes of cation diffusion facilitator proteins localized in the secretory pathway operate to activate alkaline phosphatases in vertebrate cells. (2005)
  • Author(s): Suzuki, T., Ishihara, K., Migaki, H., Nagao, M., Yamaguchi-Iwai, Y., Kambe, T.
  • Journal Title: J. Biol. Chem. 280 Pages: 30956-30962
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Involvement of heme regulatory motif in heme-mediated ubiquitination and degradation of IRP2. (2005)
  • Author(s): Ishikawa, H., Kato, M., Hori, H., Ishimori, K., Kirisako, T., Tokunaga, F., Iwai, K.
  • Journal Title: Mol Cell 19 Pages: 171-181
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Identification, expression, and assay of an oxidation-specific ubiquitin ligase, HOIL-1. (2005)
  • Author(s): Iwai, K., Ishikawa, H., Kirisako, T.
  • Journal Title: Methods Enzymol 398 Pages: 256-271
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Systematic yeast synthetic lethal and synthetic dosage lethal screens identify genes required for chromosome segregation. (2005)
  • Author(s): Measday, V., Baetz, K., Guzzo, J., Yuen, K., Kwok, T., Sheikh, B., Ding, H., Ueta, R., Hoac, T., Cheng, B., Pot, I., Tong, A., Yamaguchi-Iwai, Y., Boone, C., Hieter, P., Andrews, B.
  • Journal Title: Proc.Natl.Acad.Sci. USA 102 Pages: 13956-13961
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Zinc transporters, ZnT5, ZnT7 are required for the activation of alkaline phosphatases, zinc-requiring enzymes that are glycosylphosphatidylinositol-anchored to the cytoplasmic membrane. (2005)
  • Author(s): Suzuki, T., Ishihara, K., Migaki, H., Matsuura, W., Kohda, A., Okumura, K., Nagao, M., Yamaguchi- Iwai, Y., Kambe, T.
  • Journal Title: J.Biol.Chem. 280 Pages: 637-643
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Two different zinc transport complexes of cation diffusion facilitator proteins localized in the secretory pathway operate to activate alkaline phosphatases in vertebrate cells. (2005)
  • Author(s): Suzuki, T., Ishihara, K., Migaki, H., Nagao, M., Yamaguchi-Iwai, Y., Kambe, T.
  • Journal Title: J.Biol.Chem. 280 Pages: 30956-30962
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Involvement of heme regulatory motif in heme-mediated ubiquitination and degradation of IRP2. (2005)
  • Author(s): Ishikawa, H., Kato, M., Hori, H., Ishimori, K., Kirisako, T., Tokunaga, F., Iwai, K.
  • Journal Title: Mol.Cell. 19 Pages: 171-181
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] and assay of an oxidation-specific ubiquitin ligase, HOIL-1. (2005)
  • Author(s): Iwai, K., Ishikawa, H., Kirisako, T., Identification, expression
  • Journal Title: Methods Enzymol. 398 Pages: 256-271
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Two different zinc transport complexes of cation diffusion facilitator proteins localized in the secretory pathway operate to activate alkaline phosphates in vertebrate cells. (2005)
  • Author(s): Suzuki, T, Ishihara, K., Migaki, H., Ishihara, K., Nagao, M., Yamaguchi-Iwai, Y., Kambe, T.
  • Journal Title: J.Biol.Chem. 280 Pages: 30956-30962
• [Journal Article] Systematic yeast synthetic lethal and synthetic dosage lethal screens identify genes required for chromosome segregation. (2005)
  • Author(s): Measday, V., Baetz, K., Guzzo, J., Yuen, K., Kwok, T., Sheikh, B., Ding, H., Ueta, R., Hoac, T., Cheng, B., Pot, I., Tong, A., Yamaguchi-Iwai, Y., Boone, C., Hieter, P., Andrews, B.
  • Journal Title: Proc.Natl.Acad.Sci. 102 Pages: 13956-13961
• [Journal Article] Involvement of heme regulatory motif in heme-mediated ubiquitination and degradation of IRP2. (2005)
  • Author(s): Ishikawa, H., Kato, M., Hori, H., Ishimori, K., Kirisako, T., Tokunaga, F., Iwai, K.
  • Journal Title: Molecular Cell 19 Pages: 171-181
• [Journal Article] Identification, expression, and assay of an oxidation-specific ubiquitin ligase, HOIL-1. (2005)
  • Author(s): Iwai, K., Ishikawa, H., Kirisako, T.
  • Journal Title: Methods Enzymol. 398 Pages: 256-271
• [Journal Article] Zinc transporters, ZnT5 and ZnT7, are required for the activation of alkaline phosphatases, zinc-requiring enzymes that are glycosylphosphatidylinositol-anchored to the cytoplasmic membrane. (2005)
  • Author(s): Suzuki, T, Ishihara, K., Migaki, H., Matsuura, W., Kohda, A., Okumura, K., Nagao, M., Yamaguchi-Iwai, Y., Kambe, T.
  • Journal Title: J.Biol.Chem 280 Pages: 637-643
• [Journal Article] Overview of mammalian zinc transporter. (2004)
  • Author(s): Kambe, T., Yamaguchi-Iwai, Y., Sasaki R., Nagao, M.
  • Journal Title: Cell. Mol. Life Sci. 61 Pages: 49-68
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Overview of mammalian zinc transporter. (2004)
  • Author(s): Kambe, T., Yamaguchi-Iwai, Y., Sasaki R., Nagao, M.
  • Journal Title: Cell.Mol.Life Sci. 61 Pages: 49-68
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] 分子遺伝学アプローチによる鉄代謝機構研究の進展 (2004)
  • Author(s): 岩井 裕子
  • Journal Title: 生化学 76 Pages: 441-444
  • NAID: 10013052821
• [Journal Article] Structural basis of sugar-recognizing ubiquitin ligase. (2004)
  • Author(s): Mizushima, T., Hirao, T., Yoshida, Y., Lee, S.J., Chiba, T., Iwai, K., Yamaguchi, Y., Kato, K., Tsukihara, T., Tanaka, K.
  • Journal Title: Nat.Struct.Mol.Biol. 11 Pages: 365-370
• [Journal Article] The Ubiquitin System : From Basic Mechanisms to the Patient Bed. (2004)
  • Author(s): Ciechanover, A, Iwai, K.
  • Journal Title: IUBMB Life 56 Pages: 193-201
• [Journal Article] Structural basis for distinct roles of Lys63- and Lys48-linked polyubiquitin chains. (2004)
  • Author(s): Tenno, T., Fujiwara, K., Tochio, H., Iwai, K., Morita, E.H., Hayashi, H., Murata, S., Hiroaki, H., Sato, M., Tanaka, K., Shirakawa, M.
  • Journal Title: Genes Cells. 9 Pages: 865-875
• [Journal Article] Mechanism Underlying the Iron-Dependent Nuclear Export of the Iron-Responsive Transcription Factor Aftlp in Saccharomyces Cervisiae
  • Author(s): Ueta R., Fujiwara N., Iwai K., Yamaguchi-Iwai Y.
  • Journal Title: Molecular Biology of the Cell (In press)
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Mechanism Underlying the Iron-Dependent Nuclear Export of the Iron-Responsive Transcription Factor Aftlp in Saccharomyces Cervisiae
  • Author(s): Ueta R., Fujiwara N., Iwai K., Yamaguchi-Iwai Y.
  • Journal Title: Molecular Biology of the Cell (In press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Sequence similarlity and functional relationship among eukaryotic ZIP and CDF transporters
  • Author(s): Kambe, K., Suzuki, T., Nagao, M., Yamaguchi-Iwai, Y.
  • Journal Title: Genomics Proteomics and Bioinformatics (in press)