The research of the regulation of transcription in histidine decarboxylase gene

组氨酸脱羧酶基因转录调控的研究

• 批准号: 12670078
• 负责人: OHTSU Hiroshi
• 金额: $ 2.37万
• 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: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670078/
• 关键词: histamine; transcription; histidine decarboxylase; methylation; mast cells; transcription factor; promoter; demethylation

We have been investigating the transcriptional regulation of a gene coding histidine decarboxylase, the enzyme catalyze histamine synthesis reaction. The expression of this gene is restricted to the mast cells, basophils, neuron, gastric wall etc. The cell specific expression is controlled the methylation of the cytosine of promoter-enhancer region of this gene. We studied this phenomenon in chiefly a cell line called P815 which is a mast cell line. The following statement is the summary of the results.1) We cloned the mouse genomic DNA of HDC gene and determined the structure.2) We found that when we treated this cell line with an demethylation agent called 5-azaC, the cell newly expressed HDC gene.3) We transfected this cell line with two kinds of HDC promoter-reporter plasmids ; a plasmid without any methylation and with methylation in the HDC promoter region. The reporter activity was much weaker in the cells with methylated plasmid.4) The P815 cell line with methylated promoter-reporter construct increased the reporter activity after the stimulation with 5-aza C.The mouse HDC gene is controlled by the methylation of the promoter region which is very similar phenomenon seen in human counterparts.We constructed HDC promoter-GFP vector to check the expression of HDC gene by introducing this gene to generate the transgenic mice. These mice can be used for the transcription assay of HDC by various stimuli and the elucidation of the relation between the inducer and the transcriptional activation in vivo.

我们一直在研究编码组氨酸脱羧酶的基因的转录调控，该酶催化组胺合成反应。该基因的表达仅限于肥大细胞、嗜碱性细胞、神经元、胃壁等。该基因的细胞特异性表达受启动子增强子区胞嘧啶甲基化控制。我们主要在一种叫做P815的肥大细胞系中研究了这种现象。下面的声明是对结果的总结。1)克隆了HDC基因的小鼠基因组DNA，并确定了其结构。2)我们发现，当我们用一种叫做5-azaC的去甲基化剂处理该细胞系时，细胞新表达了HDC基因。3)用两种HDC启动子-报告质粒转染该细胞系；没有甲基化且在HDC启动子区域有甲基化的质粒。在质粒甲基化的细胞中，报告基因活性较弱。4)具有甲基化启动子-报告子结构的P815细胞系在5-aza c刺激后报告子活性增加。小鼠HDC基因受启动子区域甲基化控制，这与人类相似。我们构建HDC启动子- gfp载体，通过导入HDC基因产生转基因小鼠来检测HDC基因的表达。这些小鼠可用于各种刺激下的HDC转录测定，并阐明诱导剂与体内转录激活的关系。

项目成果

Satsuki Suzuki-Ishigaki et al.: "The mouse L-histidine decarboxylase gene : structure and transcriptional regulation by CpG methylation in the promoter region"Nucleic Acids Research. 28(14). 2627-2633 (2000)

Satsuki Suzuki-Ishigaki 等人：“小鼠 L-组氨酸脱羧酶基因：启动子区域 CpG 甲基化的结构和转录调控”核酸研究。

Satsuki Suzuki, Ishigaki et al.: "The mouse L-histidine decarboxylase gene shouter and transcriptional rognlation by CpG metheytation in the prosecutor region"Nucleic Acids Research. 28(14). 2627-2633 (2000)

Satsuki Suzuki、Ishigaki 等人：“小鼠 L-组氨酸脱羧酶基因在检察官区域通过 CpG 甲基化进行转录调控”核酸研究。

Kuramasu, A. et al.: "Identification of novel mast cell genes by serial analysis of gene expression in cord blood-derived mast cells"FEBS Letters. 498. 37-41 (2001)

Kuramasu, A. 等人：“通过对脐带血来源的肥大细胞中的基因表达进行系列分析来鉴定新型肥大细胞基因”FEBS Letters。

Ajoy Kumar Ghosh et al.: "Defective angiogenesis in the inflammation granulation tissue in histidine decarbosylase-deficient mice but not mast cell-deficient mice"J.Exp.Med.(in press). (2002)

Ajoy Kumar Ghosh 等人：“组氨酸脱碳基酶缺陷型小鼠炎症肉芽组织中的血管生成缺陷，但肥大细胞缺陷型小鼠则不然”J.Exp.Med.（出版中）。

Kuramasu, A. et al.: "Identification of novel mast cell genies by serial analysis of gene expression in cord blood-deviled mast cells"FEBS Letters. 498. 37-41 (2001)

Kuramasu, A. 等人：“通过对脐带血污染的肥大细胞中的基因表达进行系列分析来鉴定新型肥大细胞基因”FEBS Letters。