权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Thrombopoiesis regulated by a transcription factor NF-E2

转录因子 NF-E2 调节血小板生成

基本信息

批准号：
17390074
负责人：
MOTOHASHI Hozumi
金额：
$ 9.34万
依托单位：
Tohoku University (2006)University of Tsukuba (2005)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
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/grant/KAKENHI-PROJECT-17390074/
关键词：
NF-E2 megakaryocytes platelets protein complex microarray transgenic mice 少Maf群蛋白質転写制限 NF-E2 p45 胞体突起形成

项目摘要

The goal of this study was to clarify the regulatory mechanisms of thrombopoiesis through analyzing the function of a heterodimeric transcription factor NF-E2.1. Structure and function analysis of small MafA deletion mutant of MafG that lacks 40 amino acids of its C-terminus did not rescue the defective proplatelet formation of megakaryocytes from mafG-null mice when the mutant MafG was supplied from the transgene. This results clearly showed that the MafG C-terminal region is critical to the small Ma function in megakaryocytes. We investigated the proteins interacting to the MafG C-terminal region through the yeast two hybrid screening. We found that NF-E2 p45 interacts with the full length of MafG but not with MafG without the C-terminus. Interestingly, Nrf2, another member of CNC family transcription factor and a related factor of NF-E2 p45, does interact with MafG irrespective of the presence the C-terminal region. These results imply that MafG C-terminal region may contribute to t … More he selection of a heterodimeric partner molecule of MafG.2. Subcellular localization of NF-E2 p45We established transgenic mouse lines expressing Flag-tagged NF-E2 p45 or GFP-fused NF-E2 p45 in megakaryocytes. We also obtained a stable transformant megakaryocytic cell lines expressing Flag-HA-His tagged NF-E2 p45. By using these mice and cells, we are examining the subcellular distribution of NF-E2 p45.3. Target genes of NF-E2 p45We identified a novel gene clone 325 from the subtraction screening of small Maf mutant megakaryocytes and the control megakaryocytes. We disrupted the 325 gene in mice and found that the mice is healthy and fertile displaying the normal platelet count. We are planning to administer anti-platelet antibody and/or myelosupressive reagents to the mice and to see their effects on the thrombopoiesis. We also performed transcriptome analysis of NF-E2 p45-null mice and found that the factors involved in the platelet function and structures are decreased while antioxidant response genes and detoxifying enzyme genes were increased, which suggests the involvement of oxidative stress in the process of megakaryopoiesis and thrombopoiesis. Less

本研究的目的是通过分析异二聚体转录因子NF-E2.1的功能来阐明血栓形成的调控机制。对c端缺失40个氨基酸的MafA小缺失突变体的结构和功能分析表明，当从转基因中提供突变体MafG时，没有挽救mag缺失小鼠巨核细胞形成缺陷的前血小板。这一结果清楚地表明，在巨核细胞中，mag c端区域对小Ma功能至关重要。我们通过酵母双杂交筛选研究了与MafG c端区相互作用的蛋白。我们发现NF-E2 p45与全长的MafG相互作用，但不与不含c末端的MafG相互作用。有趣的是，Nrf2是CNC家族转录因子的另一个成员，也是NF-E2 p45的相关因子，无论是否存在c端区域，它都与MafG相互作用。这些结果表明，MafG c端区可能对其异二聚体伴侣分子的选择有一定的作用。我们建立了在巨核细胞中表达flag标记的NF-E2 p45或gfp融合的NF-E2 p45的转基因小鼠系。我们还获得了表达Flag-HA-His标记的NF-E2 p45的稳定转化巨核细胞系。通过使用这些小鼠和细胞，我们正在检查NF-E2 p45.3的亚细胞分布。NF-E2 p45靶基因通过Maf小突变体巨核细胞和对照巨核细胞的减法筛选，鉴定出一个新的基因克隆325。我们破坏了小鼠的325基因，发现小鼠是健康的和可生育的，显示出正常的血小板计数。我们计划给小鼠注射抗血小板抗体和/或骨髓抑制试剂，观察它们对血小板生成的影响。我们还对NF-E2 p45缺失小鼠进行了转录组分析，发现与血小板功能和结构相关的因子减少，而抗氧化反应基因和解毒酶基因增加，这表明氧化应激参与了巨核生成和血小板生成过程。少

项目成果

期刊论文数量（0）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Nrf2 Neh5 domain is differentially utilized in the transactivation of cytoprotective genes.

DOI：
10.1042/bj20061611
发表时间：
2007-06
期刊：
The Biochemical journal
影响因子：
0
作者：
Jianyong Zhang;Tomonori Hosoya;Atsushi Maruyama;K. Nishikawa;J. Maher;T. Ohta;H. Motohashi;A. Fukamizu;S. Shibahara;K. Itoh;Masayuki Yamamoto
通讯作者：
Jianyong Zhang;Tomonori Hosoya;Atsushi Maruyama;K. Nishikawa;J. Maher;T. Ohta;H. Motohashi;A. Fukamizu;S. Shibahara;K. Itoh;Masayuki Yamamoto

Constitutive expression of aryl hydrocarbon receptor in keratinocytes causes inflammatory skin lesions