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Regulation of histamine synthesis and degradation at the gene level

基因水平上组胺合成和降解的调控

基本信息

批准号：
08044230
负责人：
WATANABE Takehiko
金额：
$ 7.62万
依托单位：
Tohoku University
依托单位国家：
日本
项目类别：
Grant-in-Aid for international Scientific Research
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

项目摘要

Histidine decarboxylase (HDC) is a unique enzyme catalyzing synthesis reaction of histamine from histidine and its expression is restricted in mast cell and basophil lineage among the hematopoietic cells. We found that the methylation of promoter region of this gene suppress the transcription and those cell lineages (mast cell and basophil) have exclusively demethylated state around promoter region. Azacytidine, an demethylating reagent, produce the demethylation of promoter region of HDC gene and induce HDC expression in P815 cells, suggesting that the methylation is mechanically very important for the cell specific expression of this gene. We are going to seek for other DNase I hypersensitive sites other than the site we already found. We hope these sites contain cell specific cis-element to contribute for cell-specific demethylation which will lead the future investigation. We also proved that 74 kDa nascent HDC protein are translated in cytosol and targeted into ER to produce 53 kDa processed protein. We found that the 74 kDa protein is not only converted to 53 kDa protein but also catabolyzed viaubiquitinization and this speed of catabolism is much faster than that of the 53 kDa protein. We made HDC knockout mice by homologous recombination technique and proved that the knockout mice possess no histamine in the body (ahistaminergic mice). This mice Seemed to grow normally and were fertile. The proliferation pattern of this mice follows Mendelian law, therefore there seems to be no embryonic death. We are planning to assess the behavior and allergy using knockout mice. Once turned to catabolizing enzyme of histamine, we cloned human histamine N-methyltransferase (HMT) gene. HMT gene consists of 6 exons and resides in 2nd Chromosome q13-21. We also started to construct a plasmid for transgenic mice to produce high amount of rat HMT.

组氨酸脱羧酶(HDC)是一种独特的催化组氨酸合成组胺反应的酶，其表达仅限于造血细胞中的肥大细胞和嗜碱性粒细胞。我们发现该基因启动子区域的甲基化抑制了转录，并且那些细胞系(肥大细胞和嗜碱性粒细胞)在启动子区域周围具有专一性的去甲基化状态。去甲基化试剂氮杂胞苷在P815细胞中产生HDC基因启动子区域的去甲基化，并诱导HDC表达，提示甲基化对该基因的细胞特异性表达具有重要的机械意义。我们将寻找除了我们已经发现的部位之外的其他DNase I超敏部位。我们希望这些位点含有细胞特异的顺式元件，有助于细胞特异性去甲基化，这将引领未来的研究。我们还证明了74 kDa的新生HDC蛋白在胞浆中被翻译并靶向内质网，产生53 kDa的加工蛋白。我们发现，74 kDa的蛋白质不仅转化为53 kDa的蛋白质，而且分解代谢的病毒泛素化速度比53 kDa的蛋白质快得多。我们利用同源重组技术建立了HDC基因敲除小鼠，并证明该基因敲除小鼠体内不存在组胺(A组胺能)。这只小鼠似乎生长正常，并具有生育能力。这只小鼠的增殖模式遵循孟德尔定律，因此似乎没有胚胎死亡。我们正计划用基因敲除小鼠来评估它们的行为和过敏反应。一旦转向组胺分解酶，我们克隆了人组胺N-甲基转移酶(HMT)基因。HMT基因由6个外显子组成，位于第二染色体Q13-21。我们还开始构建转基因小鼠的表达载体，以生产大量的大鼠HMT。