Cloning and functional analysis of the mouse vasopressin/angiotensin II dual receptor

小鼠加压素/血管紧张素II双受体的克隆及功能分析

基本信息

批准号：
17590964
负责人：
IWASAKI Yasumasa
金额：
$ 2.24万
依托单位：
Kochi 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-17590964/
关键词：
vasopressin angiotensin receptor NLR receptor LRR protein NACHT アンギオテンシンレニンシグナル伝達

项目摘要

Angiotensin II/vasopressin dual receptor (AT/VPR) gene was cloned from the rat, the protein product of which has unique receptor characteristics, i.e. both angiontensin II (AII) and vasopressin (VP) act as ligands. However, the structure of the gene has not characterized yet, and the functional characteristics of the receptor remains to be clarified. In this study, we cloned the mouse AT/VPR mRNA and gene (6.5 Kb). Although the mouse and the rat AT/AVPR genes showed extremely high similarity in their nucleotide sequences, unexpectedly, the initiation codon of the rat gene was changed from Met to Leu in the mouse, resulting in extended protein product at N-terminus (179 a.a.). The mouse AT/VPR gene consisted of at least 5 exons and 4 introns by comparing the nucleotide sequence of its cDNA. Interestingly, the gene partly shared that of NACHT/Nalp6,which belongs the pathogen-recognition receptor.We then examined the tissue distribution of the AT/VPR mRNA, and found it to be expressed in the pituitary, thalamus, cerebral cortex, hippocampus, cerebellum, liver, gall bladder, adrenal gland, colon, testis, and white adipose tissue.Expression in cardiomyocytes was weak, and no expression was observed in skeletal muscle, skin, and spleen.Finally, we constructed an expression vector of the receptor and expressed it in A10 rat vascular smooth muscle cells in vitro. We fund that, at least in our experimental condition, AII did not activate any intracellular signaling pathways. On the other hand, vasopressin modestly enhanced the response of SRE-dependent transcription, whereas markedly attenuated that of NF-kB-dependent transcription, when the AT/VPR was overexpressed.We thus conclude that AT/VPR may be working not as a classical "hormone receptor" but as a pattern recognition receptor, and is modifying the NF-kB-dependent gene transcription. The functional differences between the mouse and rat AT/VPR awaits further investigation.

血管紧张素II/加压素双受体（AT/VPR）基因是从大鼠克隆的，大鼠的蛋白产物具有独特的受体特性，即Angiontensin II（AII）和加压素（VP）作为配体。但是，该基因的结构尚未表征，并且受体的功能特征仍然有待澄清。在这项研究中，我们将小鼠在/vpr mRNA和基因（6.5 kb）中克隆。尽管小鼠和AT/AVPR基因在其核苷酸序列中显示出极高的相似性，但出乎意料的是，大鼠基因的起始密码子从小鼠的MET转换为LEU，导致N末端的蛋白质产物扩展（179 A.A.）。通过比较其cDNA的核苷酸序列，小鼠AT/VPR基因由至少5个外显子和4个内含子组成。 Interestingly, the gene partly shared that of NACHT/Nalp6,which belongs the pathogen-recognition receptor.We then examined the tissue distribution of the AT/VPR mRNA, and found it to be expressed in the pituitary, thalamus, cerebral cortex, hippocampus, cerebellum, liver, gall bladder, adrenal gland, colon, testis, and white adipose tissue.Expression在心肌细胞中，在骨骼肌，皮肤和脾脏中未观察到表达。从本文中，我们在体外构建了受体的表达载体，并在体外的A10大鼠血管平滑肌细胞中表达。我们资助至少在我们的实验条件下，AII没有激活任何细胞内信号通路。另一方面，加压素适度增强了SRE依赖性转录的响应，而当AT/VPR过度表达时，我们的NF-KB依赖性转录的响应显着地减弱了。因此，我们得出的结论是，AT/VPR可能不是作为经典的“激素受体”，而是作为模式认可受体的传递，并且是NF-KB的转录。小鼠和大鼠AT/VPR之间的功能差异等待进一步研究。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

マウスバゾプレシン/アンジオテンシンデュアル受容体のクローニングと機能解析

小鼠加压素/血管紧张素双受体克隆及功能分析

DOI：
发表时间：
2007
期刊：
日本内分泌学会雑誌第80回日本内分泌学会学術総会抄録集 Vol 83,No 1(In press)
影响因子：
0
作者：
岩崎泰正;西山充;次田誠;谷口義典;岡崎瑞穂;何静;橋本浩三
通讯作者：
橋本浩三

Folia Endocrinologia Japonica

日本叶内分泌学

DOI：
发表时间：
2007
期刊：
80^<th> Japanese Endocrine Society Vol. 83, No. 1 (in press)
影响因子：
0
作者：
Yasumasa Iwasaki;Mitsuru Nishiyama;Makoto Tsugita;Yoshinori Taniguchi;Mizuho Okazaki;He Jing;Kozo Hashimoto
通讯作者：
Kozo Hashimoto

マウスァシジォテンシン・バソプレシンデュアル受容体のクローニングと機能解析

小鼠担负压素/加压素双受体的克隆及功能分析