Involvement of general transcriptional coactivator PC4 in the transcription of medaka fish intestine-specific membrane guanylyl cyclase gene (OlGC6).
Involvement of general transcriptional coactivator PC4 in the transcription of medaka fish intestine-specific membrane guanylyl cyclase gene (OlGC6).
复制标题
DOI：
10.1093/jb/mvi066
复制
发表时间：
2005-04
影响因子：
2.7
通讯作者：
M. Nakauchi;K. Yoshino;K. Yonezawa;N. Suzuki
中科院分区：
文献类型：
--
作者：
M. Nakauchi;K. Yoshino;K. Yonezawa;N. Suzuki
最近的一项研究表明，青冈鱼肠道特异性膜鸟苷酸环化酶基因OlGC6中的AGACCTTTGC核苷酸序列(-90至-81)对该基因在哺乳动物培养细胞系和青冈鱼中的转录具有重要作用。利用序列特异性DNA亲和层析，从青冈鱼肠道核提取液中纯化了一个顺式调节元件结合蛋白，并用质谱仪鉴定其为青冈鱼一般转录辅活化子PC4的同源物，我们将其命名为OlPC4。OlPC4基因在青冈鱼的胚胎和各种组织中都有表达。利用17 kDa的重组OlPC4进行了紫外光(UV)交联实验和凝胶迁移率改变分析(EMSA)，证明重组OlPC4可以替代青冈鱼肠道核提取液中的天然OlPC4。在CACO-2细胞中，将OlGC6-荧光素酶融合基因与OlPC4表达载体共转染后，OlGC6启动子被激活1.5倍。
A recent study showed that the AGACCTTTGC nucleotides sequence (between -90 and -81) contained in the cis-regulatory element in an intestine-specific membrane guanylyl cyclase gene, OlGC6, of the medaka fish, Oryzias latipes, are important for the transcription of the gene in mammalian cultured cell line and in medaka fish. Using sequence-specific DNA affinity chromatography, we purified a cis-regulatory element-binding protein from a medaka fish intestinal nuclear extract and used mass spectrometry to identify it as a medaka fish homologue of general transcriptional coactivator PC4, which we designated as OlPC4. The expression of the OlPC4 gene was detected in embryos, as well as in a large variety of tissues of adult medaka fish. Using a 17-kDa recombinant OlPC4, we carried out an ultraviolet (UV) cross-linking experiment and an electrophoretic mobility shift assay (EMSA), and demonstrated that the recombinant OlPC4 can be substituted for native OlPC4 in medaka fish intestinal nuclear extracts. In CACO-2 cells, cotransfection of the OlGC6-luciferase fusion genes with an OlPC4 expression vector resulted in 1.5-fold stimulation of the OlGC6 promoter.