Sharina I G, Krumenacker J S, Martin E, Murad F
Department of Integrative Biology and Pharmacology and the Institute of Molecular Medicine, University of Texas Medical School, 6431 Fannin, Houston, TX 77030, USA.
Proc Natl Acad Sci U S A. 2000 Sep 26;97(20):10878-83. doi: 10.1073/pnas.190331697.
The structures of the genes encoding the alpha(1) and beta(1) subunits of murine soluble guanylyl cyclase (sGC) were determined. Full-length cDNAs isolated from mouse lungs encoding the alpha(1) (2.5 kb) and beta(1) (3.3 kb) subunits are presented in this report. The alpha(1) sGC gene is approximately 26.4 kb and contains nine exons, whereas the beta(1) sGC gene spans 22 kb and consists of 14 exons. The positions of exon/intron boundaries and the sizes of introns for both genes are described. Comparison of mouse genomic organization with the Human Genome Database predicted the exon/intron boundaries of the human genes and revealed that human and mouse alpha1 and beta1 sGC genes have similar structures. Both mouse genes are localized on the third chromosome, band 3E3-F1, and are separated by a fragment that is 2% of the chromosomal length. The 5' untranscribed regions of alpha(1) and beta(1) subunit genes were subcloned into luciferase reporter constructs, and the functional analysis of promoter activity was performed in murine neuroblastoma N1E-115 cells. Our results indicate that the 5' untranscribed regions for both genes possess independent promoter activities and, together with the data on chromosomal localization, suggest independent regulation of both genes.
确定了编码小鼠可溶性鸟苷酸环化酶(sGC)α(1)和β(1)亚基的基因结构。本报告展示了从小鼠肺中分离出的编码α(1)(2.5 kb)和β(1)(3.3 kb)亚基的全长cDNA。α(1) sGC基因约为26.4 kb,包含9个外显子,而β(1) sGC基因跨度为22 kb,由14个外显子组成。描述了这两个基因的外显子/内含子边界位置和内含子大小。将小鼠基因组结构与人类基因组数据库进行比较,预测了人类基因的外显子/内含子边界,并发现人类和小鼠的α1和β1 sGC基因结构相似。两个小鼠基因均定位于第三条染色体的3E3 - F1带,且被一段占染色体长度2%的片段隔开。将α(1)和β(1)亚基基因的5'非转录区亚克隆到荧光素酶报告基因构建体中,并在小鼠神经母细胞瘤N1E - 115细胞中进行启动子活性的功能分析。我们的结果表明,这两个基因的5'非转录区均具有独立的启动子活性,结合染色体定位数据,提示这两个基因受到独立调控。