Structure of the type B human natriuretic peptide receptor gene and association of a novel microsatellite polymorphism with essential hypertension.
Structure of the type B human natriuretic peptide receptor gene and association of a novel microsatellite polymorphism with essential hypertension.
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B型人利钠肽受体基因的结构以及新型微卫星多态性与原发性高血压的关联。
DOI：
10.1161/01.res.84.5.605
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发表时间：
1999
影响因子：
20.1
通讯作者：
Y. Ozawa
中科院分区：
文献类型：
--
作者：
D. Rehemudula;T. Nakayama;M. Soma;Y. Takahashi;J. Uwabo;M. Sato;Y. Izumi;K. Kanmatsuse;Y. Ozawa
利钠肽（NP）系统可能在原发性高血压（EH）的发生发展中起重要作用。c型NP通过B型NP受体（NPR-B）扩张动脉、降低血压、抑制血管平滑肌细胞增殖。然而，人类NPR-B基因与EH的关系尚未研究，因为对该基因的基因组组织知之甚少。根据人NPR-B基因cDNA序列设计寡核苷酸引物，进行远程聚合酶链反应（PCR）。对扩增片段直接测序，确定人NPR-B基因的外显子/内含子组织。该基因全长约16.5 kbp，由22个外显子组成，内含子-外显子连接遵循GT-AG规则。采用热不对称交错pcr （TAIL-PCR）方法对其5'侧区750个碱基对进行测序。该区域包含10个潜在的Sp1结合位点，缺乏TATA盒。cDNA末端快速扩增（RACE）显示转录起始位点为- 14bp。用杂交方法鉴定了CA/GT微卫星重复序列，并将其转化为序列标记位点（sequence-tagged site， STS）。GT微卫星重复序列定位于外显子-内含子连接处下游约150 bp的内含子2。在EH患者和年龄匹配的正常血压（NT）对照中检测到两个等位基因（GT）10和（GT）11。多元logistic线性回归分析显示，NPR-B基因型与EH显著相关（优势比1.55；95%可信区间1.02 ~ 2.35）。EH组（GT）11频率为0.316 (65/206)，NT组为0.218 (44/202)，EH组与NT组间差异有统计学意义（ch2 =4.97， P=0.026）。测定了人NPR-B基因的结构组织，并鉴定出与EH相关的新的GT重复多态性。这些结果表明，EH的一个原因是该基因或密切相关的基因或区域的突变。
The natriuretic peptide (NP) system may play a crucial role in development of essential hypertension (EH). C-type NP dilates arteries and lowers blood pressure and inhibits proliferation of vascular smooth muscle cells via the type B NP receptor (NPR-B). However, the association of the human NPR-B gene with EH has not been studied, because little is known about the genomic organization of this gene. We designed oligonucleotide primers based on the cDNA sequence of the human NPR-B gene, and long-range polymerase chain reaction (PCR) was performed. The amplified fragments were sequenced directly, and the exon/intron organization of the human NPR-B gene was determined. The gene, which spans approximately 16.5 kbp, is composed of 22 exons, and the intron-exon junctions follow the GT-AG rule. Seven hundred fifty base pairs of the 5'-flanking region were sequenced using a thermal asymmetric interlaced-PCR (TAIL-PCR) method. This region contains 10 potential Sp1 binding sites and lacks a TATA box. Rapid amplification of cDNA ends (RACE) revealed the transcriptional start site at -14 bp. A CA/GT microsatellite repeat was identified with a hybridization-based method and was converted to a sequence-tagged site (STS). The GT microsatellite repeat was localized to intron 2 approximately 150 bp downstream of the exon-intron junction. Two alleles, (GT)10 and (GT)11, were detected in both EH patients and age-matched normotensive (NT) controls. Multiple logistic linear regression analysis indicated that the NPR-B genotype is associated significantly with EH (odds ratio 1.55; 95% confidence interval, 1.02 to 2.35). The (GT)11 frequency was 0.316 (65/206) for the EH group and 0.218 (44/202) for the NT group and differed significantly between the EH and NT groups (chi2=4.97, P=0.026). The structural organization of the human NPR-B gene was determined, and a novel GT repeat polymorphism, which associated with EH, was identified. These results suggest that one cause of EH is a mutation in this gene or a closely related gene or region.