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首页> 外文期刊>Molecular and Cellular Biology >Cell-type-specific function of the C-type natriuretic peptide gene promoter in rat anterior pituitary-derived cultured cell lines.
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Cell-type-specific function of the C-type natriuretic peptide gene promoter in rat anterior pituitary-derived cultured cell lines.

机译：C型利钠肽基因启动子在大鼠垂体前叶来源的培养细胞系中的细胞类型特异性功能。

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摘要

The promoter function of the human C-type natriuretic peptide (CNP) gene in various cultured cells was examined by transient transfection assays. The CNP promoter functioned very effectively in GH3 cells, which originated from the growth hormone-producing tumor of the rat anterior pituitary and somatomammotroph phenotype, but functioned much less effectively in GH1 cells, another type of rat pituitary-derived cell with a somatotroph phenotype, and rat primary cardiocytes. The CNP promoter did not function at all in other cells, including AtT20 cells of murine pituitary corticotroph origin. Functional analyses of the deleted promoters with various 5' deletion breakpoints revealed the existence of at least two negative and one positive regulatory regions. Within the positive regulatory region (positions -54 to -19), which conferred 90% of the promoter activity in GH3 cells, two equipotent GC-rich cis elements (positions -49 to -45 and -40 to -35) were identified. Both sites shared half of the promoter activity and binding properties to the nuclear protein in GH3 cells. Rat anterior pituitary tissue contained the binding protein of the identified cis element, which was identical or similar to that of GH3 cells. With Southwestern (DNA-protein) analysis, a 70-kDa specific binding protein distinct from known factors such as SP-1, AP-2, and Pit-1 was identified in the nuclear extract of GH3 cells.
机译：通过瞬时转染测定法检查了人类C型利钠肽（CNP）基因在各种培养细胞中的启动子功能。 CNP启动子在GH3细胞中非常有效地发挥作用，GH3细胞起源于大鼠垂体前叶和体细胞营养型表型的生长激素生成肿瘤，但在GH1细胞（另一种具有体养型表型的大鼠垂体来源的细胞）中的作用则差得多和大鼠原代心肌细胞。 CNP启动子在其他细胞中根本不起作用，包括鼠垂体皮质激素来源的AtT20细胞。具有各种5'缺失断点的缺失启动子的功能分析表明，存在至少两个负调控区和一个正调控区。在赋予GH3细胞90％启动子活性的正调控区域（-54至-19位）内，鉴定出两个等价的富含GC的顺式元件（-49至-45位和-40至-35位）。两个位点都共享一半的启动子活性和与GH3细胞核蛋白的结合特性。大鼠垂体前叶组织含有已鉴定的顺式元件的结合蛋白，该蛋白与GH3细胞相同或相似。通过西南蛋白质（DNA蛋白质）分析，在GH3细胞的核提取物中鉴定出了70kDa的特异性结合蛋白，该蛋白与已知因子（例如SP-1，AP-2和Pit-1）不同。

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