Novel brain specific bHLH transcription regulator, KW8

新型脑特异性 bHLH 转录调节因子 KW8

基本信息

批准号：
09640819
负责人：
MARUYAMA Kei
金额：
$ 1.98万
依托单位：
Tokyo Metropolitan Organization for Medical Research (1999)Okazaki National Research Institutes (1997-1998)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09640819/
关键词：
KW8 Neuro D Neuro D2 NDRF bHLH transcription LTP neuron-specific 転写調節因子分化因子 NeuroD KW8 basic helix-loop-helix neuron 差分転写調節

项目摘要

KW8, a NeuroD family basic helix-loop-helix protein, was cloned during the course of screening for the genes related to long term potentiation in rat hippocampal slice. It has a bHLH domain highly homologous with that of MATH-2 and NeuroD. On the other hand, the amino-acid sequence of the other regions had little homology with those of the known proteins. This protein consists of 381 amino acids and was detected only in neural tissue, indicating that it has an important role specific to neuronal activities. Its homologue NDRF/NeuroD was also reported. Its phosphorylation and spatiotemporal distribution was studied. KW8 was expressed only in brain tissues, such as the cerebral cortex, hippocampus and cerebellum. Immunohistological studies revealed that KW8 was localized in the nuclei of neurons. On immunoblotting of rat brain tissue, COS-1 cells and Neuro2A cells overexpressing KW8, this protein was detected as several diffuse bands. Alkaline phosphatase treatment reduced the molecular weights of these bands. Metabolic labeling with ィイD132ィエD1Pi in COS-1 cells confirmed that the KW8 protein was phosphorylated in vivo. Some of the physiological functions of KW8 might be regulated by this phosphorylation. In yeast, the GAL4 fusion protein containing the C-terminal region of KW8 activated transcription of the reporter gene, suggesting that KW8 had transcriptional activity.

KW8是一种神经瘤家族碱性螺旋 - 环螺旋蛋白，在筛选的过程中被克隆了与大鼠海马切片长期增强相关的基因的筛选。它具有与Math-2和Neurod高度同源的BHLH领域。另一方面，其他区域的氨基酸序列与已知蛋白质的氨基序列几乎没有同源性。该蛋白质由381个氨基酸组成，仅在神经组织中检测到，表明它的同源NDRF/Neurod也报道了。研究了其磷酸化和空间时间分布。 KW8仅在脑组织中表达，例如脑皮质，海马和小脑。免疫组织学研究表明，KW8位于神经元的核中。关于大鼠脑组织的免疫印迹，CoS-1细胞和过表达KW8的神经2A细胞，该蛋白被检测为几个弥漫带。碱性磷酸酶处理减少了这些带的分子量。在COS-1细胞中用D132 D1PI的代谢标记证实了KW8蛋白在体内被磷酸化。 KW8的某些物理功能可能受到这种磷酸化的调节。在酵母中，含有KW8的C末端区域的GAL4融合蛋白激活了报告基因的转录，这表明KW8具有转录活性。