Roles of heme in the regulation of gene expression and cell differentiation of erythroid cells

• Project/Area Number: 10480163
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Functional biochemistry
• Research Institution: Tohoku University
• Principal Investigator: HAYASHI Norio Tohoku University, Graduate School of Medicine, Professor (00004606)
• Co-Investigator(Kenkyū-buntansha): IGARASHI Kazuhiko Hiroshima University, Faculty of Medicine, Professor (00250738) ; KOBAYASHI Akira Tohoku University, Graduate School of Medicine, Assistant Professor (50292214)
• Project Period (FY): 1998 – 1999
• Project Status: Completed (Fiscal Year 1999)
• Budget Amount: ¥9,700,000 (Direct Cost: ¥9,700,000); Fiscal Year 1999: ¥3,900,000 (Direct Cost: ¥3,900,000); Fiscal Year 1998: ¥5,800,000 (Direct Cost: ¥5,800,000)
• Keywords: Heme / Transcription / Heme-oxygenase 1 / Bach1 / ヘム生合成調節 / ヘム調節モチーフ / 転写調節 / Nrf3 / ヘムオキシゲナーゼ / コプロポルフィリノーゲンオキシダーゼ / NF-E2 / MafK / 5-アミノレブリン酸合成酵素

Research Abstract

The purpose of this study is to understand the regulatory mechanisms of gene expression promoting erythroid cell differentiation. We examined the gene regulatory mechanisms of coproporphyrinogen (CPO) gene using reporter assays in cultured cells. The CPO gene is specifically active in erythroid cells but not in non-erythroid cells. We identified a novel motif CPRE, which appeared to be important for the differential expression of the gene. We also examined the function of Nrf3 and Bach1, which are members of the CNC transcription factor family. Among the members of this family, NF-E2 p45 is considered as a key transcription factor for erythroid differentiation. Molecular dissection of Nrf3 revealed a novel motif interacting with caspase 9. Biochemical analysis revealed that Bach1 associates with heme. Heme-bound Bach1 lost its DNA binding ability, suggesting that repressor activity of Bach1 is abrogated by heme. Consistently, Bach1-null mice displayed constitutive activation of heme-oxygenase 1 gene, which is a candidate target gene of Bach1. Thus, these data support the contention that Bach1 is a sensor of heme concentration and translates the heme abundance into the gene expression.

Research Products

• [Journal Article] Molecular cloning and functional characterization of a new CNC family transcription factor Nrf3. (1999)
  • Author(s): Kobayashi, A., Itoh, E., Toki, T., Takahashi, S-i, Igarashi, K., Hayashi, N., Yamamoto, M.
  • Journal Title: J. Biol. Chem. 274 Pages: 6443-6452
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Molecular cloning and functional characterization of a new CNC family transcription factor Nrf3 (1999)
  • Author(s): Kobayashi, A., Itoh, E., Toki, T., Takahashi, S-i, Igarashi, K., Hayashi, N., Yamamoto, M.
  • Journal Title: J.Biol.Chem. 274 Pages: 6443-6452
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Deficient heme and globin synthesis in ES cells lacking the erythroid-specific δ-aminolevulinate synthase gene. (1998)
  • Author(s): Harigae, H., Suwabe, N., Weinstock, P.H., Nagai, M., Fujita, H., Yamamoto, M., Sassa, S.
  • Journal Title: Blood 91 Pages: 798-805
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Alternative promoters regulate transcription of the mouse GATA-2 gene. (1998)
  • Author(s): Minegishi, N., Ohta, J., Suwabe, N., Nakauchi, H., Ishihara, H., Hayashi, N., Yamamoto, M.
  • Journal Title: J. Biol. Chem. 273 Pages: 3625-3634
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Regulation of NF-E2 activity in erythroleukemia cell differentiation. (1998)
  • Author(s): Nagai, T., Igarashi, K., Akasaka, J., Furuyama, K., Fujita, H., Hayashi, N., Yamamoto, M., Sassa, S.
  • Journal Title: J. Biol. Chem. 273 Pages: 5358-5365
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Differential regulation of mouse coproporphyrinogen oxidase geneexpression in erythroid and non-erythroid cells. (1998)
  • Author(s): Takahashi, S-i., Taketani, S., Akasaka, J-e., Kobayashi, A., Hayashi, N., Yamamoto, M., Nagai, T.
  • Journal Title: Blood 92 Pages: 3436-3444
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Deficient heme and globin synthesis in ES cells lacking the erythroid-specific d-aminolevulinate synthase gene (1998)
  • Author(s): Harigae, H., Suwabe, N., Weinstock, P.H., Nagai, M., Fujita, H., Yamamoto, M., Sassa, S.
  • Journal Title: Blood 91 Pages: 798-805
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Alternative promoters regulate transcription of the mouse GATA-2 gene (1998)
  • Author(s): Minegishi, N., Ohta, J., Suwabe, N., Nakauchi, H., Ishihara, H., Hayashi, N., Yamamoto, M.
  • Journal Title: J.Biol.Chem. 273 Pages: 3625-3634
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Regulation of NF-E2 activity in erythroleukemia cell differentiation (1998)
  • Author(s): Nagai, T., Igarashi, K., Akasaka, J., Furuyama, K., Fujita, H., Hayashi, N., Yamamoto, M., Sassa, S.
  • Journal Title: J.Biol.Chem. 273 Pages: 5358-5365
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Differential regulation of mouse coproporphyrinogen oxidase gene expression in erythroid and non-erythroid cells (1998)
  • Author(s): Takahashi, S-i., Taketani, S., Akasaka, J-e., Kobayashi, A., Hayashi, N., Yamamoto, M., Nagai, T.
  • Journal Title: Blood 92 Pages: 3436-3444
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Kobayashi Akira: "Molecular cloning and functional characterization of a new CNC family transcription factor Nrf3"J. Biol. Chem.. 274・10. 6443-6452 (1999)
• [Publications] Yoshida Chikashi: "Long range interaction of cis-DNA elements mediated by architectural transcription factor Bach1"Gene Cells. 4・11. 643-655 (1999)
• [Publications] Kobayashi Akira: "A combinatorial code for gene expression generated by transcription factor Bach2 and MAZR (MAZ-related factor) through BTB/POZ domain"Mol. Cell. Biol.. 20・5. 1733-1746 (2000)
• [Publications] Kobayashi,Akira: "Molecular cloning and functional characterization of a new CNC family transcription factor Nrf3." J.Biol.Chem.274. 6443-6452 (1999)
• [Publications] Takahashi,Shinichiro: "Differential regulation of coporoporphyrinogen oxidase gene between erythroid and nonerythroid cells." Blood. 92. 3436-3444 (1998)
• [Publications] Muto,Akihiko: "Identification of Bach2 as a B-cell-specific partner for small Maf proteins that negatively regulate the immunoglobulin heavy chain gene 3'enhancer." EMBO J.17. 5734-5743 (1998)
• [Publications] Furuyama,Kazumichi: "ALAS2 gene encodes a pyridoxine-responsive enzyme with low activity." British J.Haematol.103. 839-841 (1998)