Molecular Mechanism of Erythroid Differentiation

红系分化的分子机制

• 批准号: 07044217
• 负责人: FUJITA Hiroyoshi
• 金额: $ 7.23万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for international Scientific Research
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07044217/
• 关键词: delta-Aminolevulinate synthase; erythroid transcription factor; heme bicsynthesis; globin biosynthesis; NF-E2; p18; ヘム; 赤血球分化; 鉄芽球系貧血; ヘム代謝; δ-アミルブリン酸合成酵素; 先天性ヘム代謝障害

We have investigated the special roles of erythroid type transcription factors, such as GATA-1 and NF-E2, on erythroid cell differentiation with special reference to heme biosynthesis. When expressions of erythroid specific delta-Aminolevulinate synthase (ALAS-E) in mouse erythroleukemia (MEL) cells was disturbed by anti-sense technique, expressions of every enzymes for heme biosynthesis pathway decreaded possibly due to decline in NF-E2 (Meguro et al., 1996) . This ovservation is in good agreement with previous results in DMSO-resistant clone of MEL cells (Fujita et al., 1991) .Then, we constructed ES cells lacking ALAS-E gene. The cells expressed almost same amount of GATA-1 and NF-E2 when compared with wild type cells. The cells, however, cannot accumulate heme, benzidine positive cells, and beta-major globin mRNAs after undergoing erythroid differentiation (Haigae et al., 1998) . Thus, it is obvious that ALAS-E mediated heme biosynthesis regulates erythroid differentiation not only through transcription activity of NF-E2 (Nagai et al, 1998) but also through mature type globin synthesis during late stage of erythroid differentiation.It is also demonstrated that mice lacking p18 subunit of NF-E2 was lethal. Using testis specific promoter, GATA-1 knock down mice was also obtained. The knock down mice was also lethal, however, analyzes of fetus indicated a marked decline in hemoglobinized cells.As far as we examined nonspecific delta-aminolevulinate synthase (ALAS-N), the gene was negatively regulated via NF-KB binding sequence (Fujita, 1998) .

我们已经研究了红系类型转录因子，如加塔-1和NF-E2，对红系细胞分化的特殊作用，特别是血红素生物合成。当通过反义技术干扰小鼠红白血病（MEL）细胞中红系特异性δ-氨基乙酰丙酸合酶（ALAS-E）的表达时，血红素生物合成途径的每种酶的表达可能由于NF-E2的下降而下降（目黑等人，1996年）的报告。该排卵与MEL细胞的DMSO抗性克隆的先前结果（Fujita等人，1991）。然后，我们构建了缺乏ALAS-E基因的ES细胞。当与野生型细胞相比时，细胞表达几乎相同量的加塔-1和NF-E2。然而，细胞在经历红系分化后不能积累血红素、联苯胺阳性细胞和β-主要珠蛋白mRNA（Haigae et al.，1998年）。因此，很明显ALAS-E介导的血红素生物合成不仅通过NF-E2的转录活性调节红系分化（永井et al，1998），而且还通过红系分化晚期成熟型珠蛋白的合成调节红系分化。利用睾丸特异性启动子，获得了加塔-1基因敲除小鼠。敲除小鼠也是致死的，然而，胎儿分析表明血红蛋白化细胞显著下降，就我们检测的非特异性δ-氨基乙酰丙酸合酶（ALAS-N）而言，该基因通过NF-κ B结合序列负调控（Fujita，1998）。

项目成果

T.Nagai, H.Harigae, et al.: "5-Aminolevulinate synthase expression and hemoglobin synthesis in a human myelogenous leukemia cell lines" J.Biochem.121. 487-495 (1997)

T.Nagai、H.Harigae 等人：“人骨髓性白血病细胞系中的 5-氨基乙酰丙酸合酶表达和血红蛋白合成”J.Biochem.121。

S.Sassa, M.Kondo, et al.: "Molecular defects of coproporphyrin oxidase gene in hereditary coproporphyria." Cell.Mol.Biol.43. 59-66 (1997)

S.Sassa、M.Kondo 等人：“遗传性粪卟啉症中粪卟啉氧化酶基因的分子缺陷。”

N.Komatsu et al.: "Establishment and characterization of the thrombopoietin-dependent megakaryotic cell line,UT7/TP0." Blood. 87. 4552-4560 (1996)

N.Komatsu 等人：“血小板生成素依赖性巨核细胞系 UT7/TP0 的建立和表征。”

N.Komatsu et al.: "Establishment and characterization of the thrombopoietin-dependent megakaryotic cell line, UT7/TPO" Blood. 87. 4552-4560 (1996)

N.Komatsu 等人：“血小板生成素依赖性巨核细胞系 UT7/TPO 的建立和表征”血液。

S Sassa,H Fujita et al.: "Molecular defects of coproporphyrin oxidase gene in hereditary coproporphyria." Cell. Mol. Biol.(in press). (1997)

S Sassa、H Fujita 等人：“遗传性粪卟啉症中粪卟啉氧化酶基因的分子缺陷。”