期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：2003
卷号：100
期号：20
页码：11523-11528
DOI：10.1073/pnas.1934338100
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：Soluble guanylyl cyclase (sGC) is a cytosolic enzyme producing the intracellular messenger cyclic guanosine monophosphate (cGMP) on activation with nitric oxide (NO). sGC is an obligatory heterodimer composed of and {beta} subunits. We investigated human {beta}1 sGC transcriptional regulation in BE2 human neuroblastoma cells. The 5' upstream region of the {beta}1 sGC gene was isolated and analyzed for promoter activity by using luciferase reporter constructs. The transcriptional start site of the {beta}1 sGC gene in BE2 cells was identified. The functional significance of consensus transcriptional factor binding sites proximal to the transcriptional start site was investigated by site deletions in the 800-bp promoter fragment. The elimination of CCAAT-binding factor (CBF) and growth factor independence 1 (GFI1) binding cores significantly diminished whereas deletion of the NF1 core elevated the transcription. Electrophoretic mobility-shift assay (EMSA) and Western analysis of proteins bound to biotinated EMSA probes confirmed the interaction of GFI1, CBF, and NF1 factors with the {beta}1 sGC promoter. Treatment of BE2 cells with genistein, known to inhibit the CBF binding to DNA, significantly reduced protein levels of {beta}1 sGC by inhibiting transcription. In summary, our study represents an analysis of the human {beta}1 sGC promoter regulation in human neuroblastoma BE2 cells and identifies CBF as a critically important factor in {beta}1 sGC expression.
