Gene manipulation of heme synthetic pathway enzymes

血红素合成途径酶的基因操作

• 批准号: 10557015
• 负责人: HAYASHI Norio
• 金额: $ 6.85万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10557015/
• 关键词: Heme; Erythroid cells; Knockout mice; Transgenic mice; 5-アミノレブリン酸合成酵素; ヘム生合成; 遺伝子改変マウス; 鉄芽球性貧血; 鉄代謝; 環状鉄芽球; 赤血球分化; モデル動物

Heme biosynthesis pathway contains a battery of tissue-specific enzymes, including ALAS-E (Alas2). Our purpose is to clarify the pathogenesis of congenital diseases caused by the defective heme synthesis, such as X-linked sideroblastic anemia (XLSA), through generating an ALAS-E knockout line of mice. We generated ALAS-E knockout mouse line through gene targeting experiments. The ALAS-E-deficient mice were embryonic lethal. The cause of the lethality appeared to be the defective primitive hematopoiesis. Chimeric analysis of ALAS-E-null ES cells revealed the presence of sideroblasts that are similar to those observed in human XLSA patients. It was highly expected that the ring sideroblasts were originated from the ALAS-E-null ES cells. We then generated transgenic (TG) mouse lines expressing ALAS-E mutant molecule isolated from the human samples of XLSA patients under the regulation of GATA1-hematopoietic regulatory domain (G1HRD). When G1HRD-ALAS-Emut TG mice were crossed into ALAS-E-null background, severe phenotypes of ALAS-E-null mice were rescued only partially, suggesting that full-activity of ALAS-E is required to fully rescue the knockout phenotype. The results indicate that the ALAS-E mutant molecule is functionally impaired. These results thus demonstrate that close relationship between heme synthesis and erythropoiesis.

血红素生物合成途径包含一系列组织特异性酶，包括 ALAS-E (Alas2)。我们的目的是通过生成 ALAS-E 敲除小鼠系来阐明由血红素合成缺陷引起的先天性疾病的发病机制，例如 X 连锁铁粒幼细胞贫血 (XLSA)。我们通过基因打靶实验生成了 ALAS-E 敲除小鼠系。 ALAS-E 缺陷小鼠胚胎死亡。致命的原因似乎是原始造血功能缺陷。对 ALAS-E-null ES 细胞的嵌合分析揭示了铁粒幼细胞的存在，这与人类 XLSA 患者中观察到的相似。人们高度预期环形铁粒幼细胞起源于 ALAS-E-null ES 细胞。然后，我们在 GATA1 造血调节域 (G1HRD) 的调节下生成了表达从 XLSA 患者的人类样本中分离出的 ALAS-E 突变分子的转基因 (TG) 小鼠系。当 G1HRD-ALAS-Emut TG 小鼠与 ALAS-E-null 背景杂交时，ALAS-E-null 小鼠的严重表型仅得到部分挽救，这表明需要 ALAS-E 的完整活性才能完全挽救敲除表型。结果表明 ALAS-E 突变分子功能受损。这些结果由此证明血红素合成与红细胞生成之间的密切关系。

项目成果

Deficient heme and globin synthesis in ES cells lacking the erythroid-specific δ-aminolevulinate synthase gene.

缺乏红细胞特异性 δ-氨基乙酰丙酸合酶基因的 ES 细胞中血红素和珠蛋白合成不足。

• 发表时间: 1998
• 作者: Harigae; H.; Suwabe; N.; Weinstock; P.H.; Nagai; M.; Fujita; H.; Yamamoto; M.; Sassa; S.
• 通讯作者: S.

Furuyama,Kazumichi: "ALAS2 gene encodes a pyridoxine-responsive enzyme with low activity." British J.Haematol.103. 839-841 (1998)

Furuyama, Kazumichi：“ALAS2 基因编码一种低活性的吡哆醇反应酶。”

Differential regulation of mouse coproporphyrinogen oxidase gene expression in erythroid and non-erythroid cells

红细胞和非红细胞中小鼠粪卟啉原氧化酶基因表达的差异调节

• 发表时间: 1998
• 作者: Takahashi; S
• 通讯作者: S

Kobayashi,Akira: "Molecular cloning and functional characterization of a new CNCfamily transcription factor Nrf3." J.Biol.Chem.274. 6443-6452 (1999)

Kobayashi, Akira：“新型 CNC 家族转录因子 Nrf3 的分子克隆和功能表征。”

Regulation of NF-E2 activity in erythroleukemia cell differentiation.

红白血病细胞分化中 NF-E2 活性的调节。

• 发表时间: 1998
• 作者: Nagai; T.; Igarashi; K.; Akasaka; J.; Furuyama; K.; Fujita; H.; Hayashi; N.; Yamamoto; M.; Sassa; S.
• 通讯作者: S.