Analysis of the promoter region of human placenta-specific DSCR4 gene
Analysis of the promoter region of human placenta-specific DSCR4 gene
复制标题
DOI：
10.1016/j.bbagrm.2007.09.005
复制
发表时间：
2008-01-01
影响因子：
4.7
通讯作者：
Shimizu, Yoshiko
中科院分区：
文献类型：
--
作者：
Asai, Satoko;Yamaki, Akiko;Shimizu, Yoshiko
DSCR 4基因位于人类21号染色体q22.2带，编码一个118个氨基酸的蛋白质。DSCR 4的表达仅限于人胎盘和胎盘绒毛膜癌细胞系BeWo和JEG 3。利用人胎盘RNA的5 '-RACE方法表明，主要的转录起始位点在起始密码子上游93 nt（nt-93）。使用与荧光素酶报告基因融合的5 ′-侧翼区的一系列缺失构建体的转染测定鉴定了三个阳性区域nt-2200至-2088，nt-2064至-1924，nt-810至-632和两个阴性区域nt - 1923至-1740，nt-631至-425。计算机分析预测在这些区域中存在几个顺式元件，并且使用共有序列的各种突变体的启动子测定鉴定了OLF-1和E47的两个不同的顺式元件。使用DSCR 4表达细胞的提取物的电泳迁移率变动分析（EMSA）证实了某些蛋白因子与这些顺式元件的结合。事实上，在DSCR 4表达细胞系和人胎盘中检测到OLF-1样转录因子EBF-3和EBF-4。基于这些数据，我们推测DSCR 4基因的转录受OLF-1样转录因子的正调控，受E47样转录因子的负调控。(c)2007 Elsevier B. V.保留所有权利。
The gene DSCR4 locates in the band q22.2 of human chromosome 21 and encodes a protein of 118 amino acids. Expression of DSCR4 is restricted to human placenta and placental choriocarcinoma cell lines BeWo and JEG3. The 5'-RACE method using RNA from human placenta indicated the major transcription start site at 93 nt upstream (nt -93) of the initiation codon. Transfection assay using a series of deletion constructs of the 5'-flanking region fused to the luciferase reporter gene identified three positive regions nt -2200 to -2088, nt -2064 to - 1924, nt -810 to -632 and two negative regions nt - 1923 to - 1740, nt -631 to -425. The computer analysis predicted the presence of several cis-elements in these regions and the promoter assay using various mutants of consensus sequence identified two distinct cis-elements for OLF-1 and E47. The electrophoretic mobility shift assay (EMSA) using the extracts of DSCR4-expressing cells confirmed the binding of certain protein factors to these cis-elements. In fact, OLF-1-like transcription factor, EBF-3 and EBF-4 were detected in the DSCR4-expressing cell lines and human placenta. Based on these data, we postulated that transcription of DSCR4 gene is regulated positively by binding of OLF-1-like transcription factor and negatively by binding of E47-like transcription factor. (c) 2007 Elsevier B.V. All rights reserved.