Expression of delta-aminolevulinate synthase durin erythroid differentiation

红系分化过程中δ-氨基乙酰丙酸合酶的表达

• 批准号: 05670136
• 负责人: FUJITA Hiroyoshi
• 金额: $ 1.34万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05670136/
• 关键词: Delta-aminolevulinate synthase; Erythroid differentiation; Mouse erythroleukemia cells; Heme; Positive feedback regulation; NF-E2; δ-アミルブリン酸合成酵素; ヘム代謝; 赤血球系転写因子; β-グロビン; ヘム代謝調節

Mouse erythroleukemia (MEL) cells undergo erythroid differentiation after treatment of dimethylsulfoxide (DMSO).Recently, we established DR cell, a clone of MEL cell, which could not undergo erythroid differentiation after DMSO treatment. Analysis revealed a deficiency of erythroid-specific delta-aminolevulinate synthase (ALAS-E) in DR cells. In addition, gene activation of delta-aminolevulinate dehydratase, porphobilonogen deaminase, and uroporphyrinogen decarboxylase-after DMSO treatment were reduced to approx. 1/3 of those in wild type cells.In the present study we evidenced that deficiency of heme in DR cells resulted in reduced concentrations of minas encoding ferrochelatase and beta-globin. Further studies suggest that insufficient supply of heme inhibits the gene activation of NF-E2. It is, therefore, suggested that NF-E2 might regulate heme biosynthesis in erythroid cells in a positive feedbach manner.Then, we have developed MEL cell lines, whose ALAS-E levels were declined by antisense RNA of the enzyme. Antisense RNA expression caused reductions not only in ALAS-E expression but also in the other heme pathway enzymes. These observations are in good agreement with those observed in DR cells. Furthermore, expression of ALAS-E antisense RNA also reduced NF-E2 mRNA.Thus, it is highly probable that NF-E2 plays one of the significant roles on positive feedback regulation of heme in erythroid cells.

小鼠红白血病（MEL）细胞经二甲基亚砜（DMSO）处理后可向红系分化，我们建立了MEL细胞的克隆--DR细胞，该细胞经DMSO处理后不能向红系分化。分析显示DR细胞中红系特异性δ-氨基乙酰丙酸合酶（ALAS-E）缺乏。此外，在DMSO处理后，δ-氨基乙酰丙酸酯酶、胆色素原脱氨酶和尿卟啉原脱羧酶的基因活化降低至约100%。在本研究中，我们证明了在DR细胞中血红素的缺乏导致编码亚铁螯合酶和β-珠蛋白的minas浓度降低。进一步的研究表明，血红素供应不足会抑制NF-E2的基因激活。因此，我们认为NF-E2可能以正反馈的方式调节红系细胞血红素的合成。反义RNA表达不仅导致ALAS-E表达减少，还导致其他血红素途径酶减少。这些观察结果与在DR细胞中观察到的结果非常一致。ALAS-E反义RNA的表达也降低了NF-E2 mRNA的表达，因此，NF-E2很可能在红系细胞血红素的正反馈调节中起重要作用。

项目成果

H.Fujita et al.: "Regulation of Heme Protein Synthesis" AlphaMed Press, 139 (1994)

H.Fujita 等人：“血红素蛋白合成的调节”AlphaMed Press，139 (1994)

S.Taketani & H.Fujita: "The ferrochelatase gene structure and molecular defects associated with erythropoietic protoporphyria." J.Bioenerg.Biomembr.(in press). (1995)

竹谷S

N.Komatsu and H.Fujita: "Induced megakaryocytic maturation of the human leukemia cell line,UT-7,results in down-modulation of erythropoietin receptor gene" Cancer Res.53. 1156-1161 (1993)

N.Komatsu 和 H.Fujita：“诱导人类白血病细胞系 UT-7 巨核细胞成熟，导致促红细胞生成素受体基因下调”Cancer Res.53。

N.Komatsu, M.Yamamoto, H.Fujita, A.Niwa, K.Hatake, T.Endo, H.Okano, T.Katsube, Y.Fukumaki, S.Sassa & Y.Miura: "Establishment and characterization of an erythropoietin-dependent subline, UT-7/Epo derived from human leukemia cell line, UT-7." Blood. 82. 456

N.小松、M.Yamamoto、H.Fujita、A.Niwa、K.Hatake、T.Endo、H.Okano、T.Katsube、Y.Fukumaki、S.Sassa

S.Sassa & H.Fujita: "delta-Aminolevulinate synthase : its properties and regulation" Regulation of Heme Biosynthesis by Drugs, Hormones and Environmental Chemicals. (in press). (1995)

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