Biglycan Gene Expression in the Human Leiomyosarcoma Cell Line SK-UT-1
Biglycan Gene Expression in the Human Leiomyosarcoma Cell Line SK-UT-1
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双糖链蛋白聚糖基因在人平滑肌肉瘤细胞系 SK-UT-1 中的表达
DOI：
--
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发表时间：
1998
影响因子：
4.8
通讯作者：
H. Kalthoff
中科院分区：
文献类型：
--
作者：
H. Ungefroren;B. Gellersen;N. Krull;H. Kalthoff
在这项研究中，我们证明了编码小亮氨酸蛋白多糖的基因在人肌组织和人平滑肌肉瘤细胞系SK-UT-1中表达。用forskolin或8-bromo-cAMP处理SK-UT-1细胞可显著增加biglycan mRNA的表达，并通过biglycan启动子-荧光素酶报告融合基因的瞬时转染实验表明，这种作用是转录性的。在SK-UT-1细胞中，cAMP介导的对转染后的biglycan启动子的诱导被一种特异性蛋白激酶a抑制剂的共表达所消除，并被蛋白激酶a的催化亚基（Cβ）的过表达所模拟。通过5 '缺失分析，cAMP的部分反应定位于biglycan启动子残基- 78至- 46的片段。该区域对异源启动子具有很强的cAMP响应性。电泳迁移位移和抗体超位移分析鉴定出两种特异性复合物，它们分别含有与普遍存在的转录因子Sp1和Sp3抗原性相关的核蛋白。这些蛋白的结合位点被定位到biglycan启动子中从−59到−49的一个富含ct的序列。该序列的突变消除了复合物的形成，并显著降低了转染的报告基因的基础启动子和camp依赖启动子的活性。利用重组Sp1进行的体外结合研究表明，与CT元件结合的核因子不是Sp1，而是Sp1样蛋白。SK-UT-1核蛋白的Western blot分析证实了Sp3、Sp1和与Sp1抗体发生交叉反应的核蛋白的表达，但根据它们的分子量不是Sp1。这些结果表明，SK-UT-1细胞中所有camp依赖性转录以及一些基础的biglycan转录都是通过活化的蛋白激酶A介导的，并且这两种功能都是通过sp1样/Sp3因子与biglycan启动子中−59位CT元件的相互作用在启动子水平上赋予的。
In this study we demonstrate that the gene encoding the small leucine-rich proteoglycan biglycan is expressed in human myometrial tissue and in the human leiomyosarcoma cell line SK-UT-1. Treatment of SK-UT-1 cells with forskolin or 8-bromo-cAMP strongly increased biglycan mRNA and this effect was transcriptional as shown by transient transfection experiments with biglycan promoter-luciferase reporter fusion genes. The cAMP-mediated induction of the transfected biglycan promoter in SK-UT-1 cells was abolished by coexpression of a specific protein kinase A inhibitor, and was mimicked by overexpression of the catalytic subunit (Cβ) of protein kinase A. By 5′ deletion analysis, part of the cAMP response was localized to the segment from residues −78 to −46 of the biglycan promoter. This region conferred strong cAMP responsiveness to a heterologous promoter. Electrophoretic mobility shift and antibody supershift assays identified two specific complexes that contained nuclear proteins antigenically related to the ubiquitous transcription factors Sp1 and Sp3, respectively. The binding site of these proteins was mapped to a CT-rich sequence extending from −59 to −49 in the biglycan promoter. Mutating this sequence eliminated complex formation and markedly reduced basal and cAMP-dependent promoter activity of transfected reporter genes. In vitro binding studies using recombinant Sp1 revealed that the nuclear factor binding to the CT element was not Sp1 but a Sp1-like protein(s). Western blot analysis of SK-UT-1 nuclear proteins confirmed expression of Sp3, Sp1 and nuclear proteins that crossreacted with Sp1 antibody but according to their molecular weight were not Sp1. These results indicate that all cAMP-dependent as well as some basal biglycan transcription in SK-UT-1 cells is mediated through activated protein kinase A and that both functions are conferred at the promoter level through the interaction of Sp1-like/Sp3 factors with the CT element at −59 in the biglycan promoter.
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影响因子：
56.9
作者：
WANG, CY;PETRYNIAK, B;LEIDEN, JM
通讯作者：
LEIDEN, JM
cAMP 抗性中国仓鼠卵巢细胞系中催化亚基 mRNA 水平降低并改变催化亚基 mRNA 结构。
DOI：
--
发表时间：
1991
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Howard,P;Day,KH;Kim,KE;Richardson,J;Thomas,J;Abraham,I;Fleischmann,RD;Gottesman,MM;Maurer,RA
通讯作者：
Maurer,RA
影响因子：
4.1
作者：
HILDEBRAND, A;ROMARIS, M;RUOSLAHTI, E
通讯作者：
RUOSLAHTI, E
cAMP 依赖性蛋白激酶催化亚基的两种亚型均可激活催乳素基因的转录。
DOI：
--
发表时间：
1989
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Maurer,RA
通讯作者：
Maurer,RA
影响因子：
14.9
作者：
Kennett, SB;Udvadia, AJ;Horowitz, JM
通讯作者：
Horowitz, JM