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Regulation of erythroid differentiation by trnscription factors and heme

Research Project

Project/Area Number 08458188
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Functional biochemistry
Research InstitutionTohoku University

Principal Investigator

HAYASHI Norio  Tohoku University, School of Medicine, Professor, 医学部, 教授 (00004606)

Co-Investigator(Kenkyū-buntansha) NAGAI Tadashi  Tohoku University, School of Medicine, Assistant Professor, 医学部, 助手 (40237483)
MUNAKATA Hiroshi  Tohoku University, School of Medicine, Associate Professor, 医学部, 助教授 (90111294)
古山 和道  東北大学, 医学部, 助手 (80280874)
Project Period (FY) 1996 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥8,300,000 (Direct Cost: ¥8,300,000)
Fiscal Year 1997: ¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 1996: ¥5,000,000 (Direct Cost: ¥5,000,000)
KeywordsNF-E2 / GATA-1 / MafK / Bach1 / Bach2 / heme regulatory motif / 5-aminolevulinate synthase / coproporphyrinogen oxidase / p45 / Bach因子 / メロンボポエチン / M-TAT細胞
Research Abstract

1) We showed that dimethyl sulfoxide increased the eryThroid transcription factor NF-E2' DNA complex formation in the wild -type MEL cells, in part by the induction of p45 gene expression, and that both DNA binding and transactivation activity of NF-E2 are regulated by Ser/Thr phosphorylation such
as Ras siganling cascade. An Mafk homodimer may suppress transcription not only by competition for the DNA binding site, but also by directly inhibiting transcription.
2) Using a yeast two-hybrid screen with a GAL4-MafK fusion protein, we identified two novel bZip transcription factors, Bach1 and Bach2, as heterodimerization partners of MafK, In addition to a CNC-type bZip domain, these Bach proteins possess another protein interaction motif, BTB domain. Expression of Bach1 appears ubiquitous, but that of Bach2 is restricted to monocytes and neuronal cells. Bach proteins bind in vitro to NF-E2 binding sites by forming heterodimers with MafK.Bach1 and Bach2 function as transcription repressors i … More n transfection assays using fibroblast cells, but they function as a transcription activator and repressor, respectively, in cultured erythroid cells.
3) A short amino acid sequence, the heme regulatory motif (HRM), has been shown to be involved in the hemin inhibition of protein transport into mitochondria in vitro. To elucidate the role of HRM in the home regulation of 5-aminolevulinate synthase (ALAS) transport in vivo, we constructed a series of mutants of the non-specific isoform of ALAS (ALAS-N) in which the specific cysteine residues within the HRMs were converted to serines. Wild-type and mutant enzymes were expressed in QT6 fibroblasts through transient transfection, followed by analyses of the mitochondrial import of the enzymes. The home inhibition, which was observed in the wild-type ALAS-N, abolished completely when all the three HRMs in the enzyme were mutated, indicating that the FIRM is actually required for the heme regulation of ALAS-N transport within the cells. In contrast, exogenous hemin did not affect the import of the erythroid ALAS (ALAS-E) under the comparable experimental conditions.
4) We analyzed ALAS-E gene (ALAS2) of Japanese males with pyridoxine-refractory X-l inked sideroblastic anemia (XLSA) and identified an ALAS2 mutation (D19OV). The mutation did not affect the enzyme activity in vitro. However, the amount of the enzyme in bone marrow cells were found to be reduced to approximately 5% of the normal control. Additionally, accumulation of aberrantly processed proteins, the sizes of which were larger than that of mature ALAS-E, was found in mitochondria in transient transfection analyses. These larger products were reproducibly detected in assays combining in vitro transcription/translation and precursor import into isolated mitochondria. These results indicate that the mutation causing pyridoxine-refractory XLSA may affect the processing of ALAS-E precursor, thus provoking instability of the ALAS-E protein. Less

Report

(3 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • Research Products

    (24 results)

All Other

All Publications (24 results)

  • [Publications] Nagai,Tadashi: "Regulation of NF-E2 activity in erythroleukemia cell differentiation." J.Biol.Chem.273. 5358-5365 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Itoh,Ken: "An Nrf2 small Maf heterodimer mediates the induction of phase II detoxifying enzyme genes through antioxidant response elements." Biochem.Biophys.Res.Commun.236. 313-322 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Furuyama,Kazumichi: "Pyridoxine refractory X-linked sideroblastic anemia caused by a point mutation in the erythroid 5-aminolevulinate synthase gene." Blood. 90. 822-830 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Nagai,Mayumi: "Novel regulation of δ-aminolevulinate synthase in the rat Harderian gland." Biochem.Pharmacol.53. 643-650 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Nagai,Tadashi: "5-aminolevulinate synthase expression and hemoglobin synthesis in a human myelogenous leukemia cell line." J.Biochem.(Tokyo). 121. 487-495 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Oyake,Tatsuya: "Bach proteins belong to a novel family of BTB-basic leucine zipper transcription factors that interact with MafK and regulate transcription through the NF-E2 site." Mol.Cell.Biol.16. 6083-6095 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Nagai, T., Igarashi, K., Akasaka, J., Furuyama, K., Fujita, H., Hayashi, N., Yamamoto, M., Sassa, S.: "Regulation of NF-E2 activity in erythroleukemia cell differentiation." J.Biol.Chem.273. 5358-5365 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Itoh, K., Chiba, T., Takahashi, S., Ishii, T., Igarashi, K., Katoh, Y., Oyake, T., Hayashi, N., Satoh, K., Hatayama, I., Yamamoto, M., Nabeshima, Y.: "An Nrf2 small Maf heterodimer mediates the induction of phase II detoxifying enzyme genes through antioxidant response elements." Biochem.Biophys.Res.Commun.236. 313-322 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Furuyama, K., Fujita, H., Nagai, T., Yomogida, K., Munakata, H., Kondo, M., Kimura, A., Kuramoto, A., Hayashi, N., Yamamoto, M.: "Pyridoxine refractory X-linked sideroblastic anemia caused by a point mutation in the erythroid 5-aminolevulinate synthase gene." Blood. 90. 822-830 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Nagai, M., Nagai, T., Yamamoto, M., Goto, K., Bishop, T.R., Hayashi, N., Kondo, H., Seyama, Y., Kano, K., Fujita, H., Sassa, S.: "Novel regulation of d-aminolevulinate synthase in the rat Harderian gland." Biochem.Pharmacol.53. 643-650 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Nagai, T., Harigae, H., Furuyama, K., Munakata, H., Hayashi, N., Endo, K., Sassa, S., Yamamoto, M.: "5-aminolevulinate synthase expression and hemoglobin synthesis in a human myelogenous leukemia cell line." J.Biochem.(Tokyo). 121. 487-495 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Oyake, T., Itoh, K., Motohashi, H., Hayashi, N., Hoshino, H., Nishizawa, M., Yamamoto, M., Igarashi, K.: "Bach proteins belong to a novel family of BTB-basic leucine zipper transcription factors that interact with MafK and regulate transcription through the NF-E2 site." Mol.Cell.Biol.16. 6083-6095 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Nagai, Tadashi: "Regulation of NF-E2 astivity in erythroleukemia call differetiation." J.Biol.Chem.273・9. 5358-5365 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Itoh, Ken: "An Nrf2 small Maf heterodimer mediates the induction of phase ll detoxfying enzyme genes through antioxidant response elemnts." Biochem.Biophys.Res.Commun.236・2. 313-322 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Takahashi, Satoru: "Arrest in primitive erythroid cell development caused by promoter-specific disruption of the GATA-1 gene." J.Biol.Chem.272・19. 12611-12615 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Onodera, Ko: "GATA-1 transcription is controlled by distinct regulatory mechanisms during primitive and definitive erythropoiesis." Proc.Natl.Acad.Sci.USA. 94・9. 4487-4492 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Nagai, Mayumi: "Novel regulation of delta-aminolevinate synthase in the rat Hardeian gland." Biochem.Pharmacol.53・5. 643-650 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Nagai, Tadashi: "5-aminolevulinate synthats expression and hemoglobin synthesis in a human myelogenous leukemia cell line." J.Biochem.(Tokyo). 121・3. 487-495 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Oyake, Tatsuya: "Bach proteins belong to a novel family of BTB-basic leucine zipper transcription factor that interact with MafK and regulate transcription through the NF-E2 site" Mol. Cell. Biol.16・11. 6083-6095 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Onodera, Ko: "Comserved structure, regulatory elements, and transcriptional regulation from the GATA-1 gene testis promoter" J. Biochem.121・2. 251-263 (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Nagai, Tadashi: "5-Aminolevulinate synthase expression and hemoglobin synthesis in a human myelogenous leukemia cell line" J. Biochem.121・3. 487-495 (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Minegishi, Naoko: "Expression of GATA transcription factors in myelogenous and lymphoblastic leukemia cells" Int. J. Hematol.(印刷中). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Nagai, Mayumi: "Novel regulation of δ-amonolevulinate synthase in the rat Harderian gland" Biochem. Pharmacol.(印刷中). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Furuyama, Kazumichi: "Pyridoxine refractory X-linked sideroblastic anemia caused by a point mutation in the erythroid 5-aminolevulinate synthase gene" Blood. (印刷中). (1997)

    • Related Report
      1996 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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