Multiple mechanisms for the regulation of heme biosynthesis and catabolism.

调节血红素生物合成和分解代谢的多种机制。

基本信息

批准号：
17590262
负责人：
FURUYAMA Kazumichi
金额：
$ 2.24万
依托单位：
Tohoku University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17590262/
关键词：
heme aminolevulinate synthase heme oxygenase RNA interference アミルブリン酸合成酵素

项目摘要

Heme is a prosthetic group of several enzymes, and essential for aerobic organisms. Since excess amount of heme in the cells increases reactive oxygen species, cellular heme level is strictly regulated under the balance of its biosynthesis and catabolism. To know how heme biosynthesis and catabolism is regulated by the mechanisms through protein-protein interaction, we have performed following experiments 1. To know what kind of proteins are involved in the regulation of heme biosynthesis and catabolism, we have screened human cDNA libraries using non-specific aminolevulinate synthase, erythroid-specific aminolevulinate synthase (ALAS2), heme oxygenase 1 or heme oxygenase 2 (HO-2) as a bait protein. As a result, we have found that one of the components of 20S proteasome could associate with HO-2 protein. Since over-expression of this protein reduced HO-2 protein expression level, this protein should be involved in the regulation of protein degradation of HO-2. 2. Using RNA interference … More (RNAi) technology, we have tried to establish the model cells for sideroblastic anemia. First, we have prepared effective small interfering RNA (siRNA) for suppression of ALAS2 expression, then, prepared expression vector, which express effective short-hairpin RNA (shRNA). This vector has been introduced into YN-1 cells, which is able to produce hemoglobin in the cells after the stimulation by transforming growth factor beta 1, and these cells were incubated with G418 to select the cells which constitutively express shRNA against ALAS2. YN-1 cells that express ALAS2 at low level were selected, and named as YN1-ALAS21ow cells. However, ringed sideroblast, which is the hallmark of the sideroblastic anemia, was not observed even after the erythroid differentiation by TGF-betal. We have confirmed that YN1-ALAS21ow cells expressed lower level of ALAS2 mRNA than control cells, YN1-ALAS21ow cells might expressed enough level of ALAS2 for protecting the cells from iron accumulation in mitochondria. Less

血红素是多种酶的辅基，对需氧生物至关重要。由于细胞中过量的血红素会增加活性氧，因此细胞血红素水平在其生物合成和分解代谢的平衡下受到严格调节。血红素的生物合成是通过蛋白质-蛋白质相互作用的机制来调节的，我们进行了以下实验1、了解哪些蛋白质参与了血红素生物合成的调控。分解代谢，我们使用非特异性氨基乙酰丙酸合酶、红细胞特异性氨基乙酰丙酸合酶（ALAS2）、血红素加氧酶1或血红素加氧酶2（HO-2）作为诱饵蛋白筛选了人类cDNA文库，结果，我们发现了一种。 20S蛋白酶体的成分可能与HO-2蛋白相关，由于该蛋白的过度表达降低了HO-2蛋白的表达水平，因此该蛋白应该参与了HO-2蛋白的表达。 2.利用RNA干扰（RNAi）技术，我们尝试建立铁粒幼细胞贫血的模型细胞，首先，我们制备了有效的小干扰RNA（siRNA）来抑制ALAS2的表达。然后，制备表达有效短发夹RNA(shRNA)的表达载体，将该载体导入YN-1细胞中，经转化生长因子β1刺激后，细胞内能够产生血红蛋白。将这些细胞与G418一起孵育以选择组成型表达针对ALAS2的shRNA的细胞，选择低水平表达ALAS2的YN-1细胞，并将其命名为YN1-ALAS21ow细胞，这是铁粒幼细胞的标志。即使在通过 TGF-β 进行红系分化后也没有观察到贫血。我们已经证实 YN1-ALAS21ow 细胞表达较低水平的。与对照细胞相比，YN1-ALAS21ow 细胞的 ALAS2 mRNA 可能表达足够水平的 ALAS2，以保护细胞免受线粒体中铁积累的影响。

项目成果

期刊论文数量（18）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Aig452 substitution of the erythroid-specific 5-aminolaevulinate synthase, a hot spot mutation in X-linked sideroblastic anaemia, does not itself affect enzyme activity.

红系特异性 5-氨基乙酰丙酸合酶的 Aig452 取代是 X 连锁铁粒幼细胞贫血的热点突变，其本身并不影响酶活性。

DOI：
发表时间：
2006
期刊：
European Journal of Haematology 76巻・1号
影响因子：
0
作者：
Sudo;K.;Furuyama K.
通讯作者：
Furuyama K.

Identification of adipocyte differentiation-related regulatory element for adrenomedullin gene repression (ADRE-AR) in 3T3-Ll cells.

3T3-L1细胞中肾上腺髓质素基因抑制(ADRE-AR)的脂肪细胞分化相关调节元件的鉴定。

DOI：
发表时间：
2006
期刊：
Peptides 27巻・6号
影响因子：
0
作者：
Ding Y.;et al.;Nagata K;Yutaka Hasegawa;Li Y.
通讯作者：
Li Y.

Identification of adipocyte differentiation-related regulatory element for adrenomedullin gene repression (ADRE-AR) in 3T3-L1 cells.

鉴定 3T3-L1 细胞中肾上腺髓质素基因抑制 (ADRE-AR) 的脂肪细胞分化相关调节元件。

DOI：
发表时间：
2006
期刊：
Peptides Journal. 27巻・6号
影响因子：
0
作者：
Aiba;Y.;Li Y.
通讯作者：
Li Y.

Down-regulation of heme oxygenase-2 is associated with the increased expression of heme oxygenase-1 in human cell lines

DOI：
10.1111/j.1742-4658.2006.05526.x
发表时间：
2006-12-01
期刊：
FEBS JOURNAL
影响因子：
5.4
作者：
Ding, Yuanying;Zhang, Yong Z.;Shibahara, Shigeki
通讯作者：
Shibahara, Shigeki

Hypoxemia and attenuated hypoxic ventilatoiy responses in mice lacking heme oxygenase-2 : evidence for a novel role of heme oxygenase-2 as an oxygen sensor.

缺乏血红素加氧酶 2 的小鼠的低氧血症和减弱的缺氧通气反应：血红素加氧酶 2 作为氧传感器的新作用的证据。

DOI：
发表时间：
2006
期刊：
Advances in experimental medicine and biology 580巻
影响因子：
0
作者：
Mizuta E;Miake J;Yano S;Furuichi H;Manabe K;Sasaki N;Igawa O;Hoshikawa Y;Shigemasa C;Nanba E;Ninomiya H;Hidaka K;Morisaki T;Tajima F;Hisatome I;Zhang Y.
通讯作者：
Zhang Y.