Molecular biology of vasoactive substances

血管活性物质的分子生物学

基本信息

批准号：
06044129
负责人：
NAKAO Kazuwa
金额：
$ 2.82万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for international Scientific Research
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

项目摘要

Brain natriuretic peptide (BNP) mRNA increased and reached a maximal level within 1 h in a model of cardiac hypertrophy using cultured neonatal ratventricular cardiocytes, which was completely diminished by a transcriptional inhibitor. The promotor activity of the cloned 5'-flanking region of human BNP gene was reduced to 30% when the CT-rich sequences (-1288 to -1095) were deleted. The BNP gene was assigned to human chromosome 1. Two novel transcripts of the human endothelin-A receptor (ET-AR) gene contained deletions of 199 bp and 327 bp were demonstrated using RT-PCR.The deleted sequences corresponded to exon 4 and exons 3 and 4, respectively, indicating these ET-AR transcripts result from alternative RNA splicing. The putative negative regulatory region was located between -453 and -225 of mouse angiotensin II type 2 (AT_2) receptor gene, in which the interferon regulatory factor (IRF) binding motif was identified. IRF-2 attenuated the AT2 receptor expression in both growing and confluent R3T3 cells, whereas IRF-1 enhanced AT2 receptor expression in the confluent cells only. The human prostacyclin receptor gene spanned approximately 7.0 kb and was composed of three exons, which was assigned to chromosome 19. The transcription initiation sites were mapped 870-872 bp upstream to the ATG start codon. The 1.2-kb 5'-flanking region lacked conventional TATA and CCAAT boxes, but it contained several cis-acting regulatory elements including an inverted CCAAT box and two copies of SP-1 binding sites.

在心脏肥大模型中，使用培养的新生儿肾上腺心脏心形成细胞，在1小时内，脑发脂性肽（BNP）mRNA在1小时内增加并达到最大水平，这被转录抑制剂完全降低。当删除了富含CT的序列（-1288至-1095）时，人BNP基因的克隆5'Fancing区域的启动子活性降低至30％。 BNP基因被分配到人类染色体1。使用RT-PCR证明了人内皮素A受体（ET-AR）基因的两个新型转录本，其中包含199 bp和327 bp的缺失。 3和4分别表明这些ET-AR转录本是由替代RNA剪接引起的。假定的负调节区域位于小鼠血管紧张素II型2（AT_2）受体基因的-453和-225之间，其中鉴定了干扰素调节因子（IRF）结合基序。 IRF-2减弱了生长和汇合R3T3细胞中的AT2受体表达，而IRF-1仅在汇合细胞中增强了AT2受体表达。人类前列环素受体基因跨越约7.0 kb，由三个外显子组成，该外显子被分配给19号染色体。转录起始位点的映射为870-872 bp，上游到ATG起始密码子上游。 1.2-kb 5'乘区域缺乏常规的tata和CCAAT盒，但它包含了几个顺式作用调节元件，包括倒置CCAAT盒和两个SP-1结合位点的副本。