Identification of Two Regulatory Elements within the Promoter Region of the Mouse Connexin 43 Gene (*)
Identification of Two Regulatory Elements within the Promoter Region of the Mouse Connexin 43 Gene (*)
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小鼠连接蛋白 43 基因启动子区内两个调控元件的鉴定 (*)
DOI：
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发表时间：
1995
影响因子：
4.8
通讯作者：
S. Lye
中科院分区：
文献类型：
--
作者：
Zhi;D. Lefebvre;X. Bai;A. Reaume;J. Rossant;S. Lye
为了确定连接蛋白43基因（cx43）在子宫肌层细胞中表达所需的最小序列，我们生成了转录起始位点上游延伸1686个碱基对和下游延伸162个碱基对的片段的5′缺失构建体，并确定了它们在转染的子宫肌层细胞系中驱动氯霉素乙酰转移酶报告基因表达的能力。我们的研究揭示了该片段中的两个顺式作用调节元件。从-102到-92的区域的缺失导致启动子活性增加超过10倍，表明存在阻遏元件。从-72到-62的区域的缺失导致类似程度的启动子活性的降低，这意味着正元件的存在。电泳迁移率变动分析表明，来自这两个小区域的合成寡核苷酸可以分别与从子宫肌层细胞制备的核蛋白结合，并且将三个和两个碱基取代引入这些寡聚体中的每一个足以消除它们的蛋白结合能力。这些相同的突变，当掺入氯霉素乙酰转移酶构建体中时，减弱了阴性和阳性元件的调节功能，并且在已知表达cx43基因的几种组织中发现了与这些功能元件结合的蛋白质。
To define the minimal sequences required for expression of the connexin 43 gene (cx43) in myometrial cells, we generated 5′ deletion constructs of a fragment extending 1686 base pairs upstream and 162 base pairs downstream of the transcription start site and determined their ability to drive expression of the chloramphenicol acetyltransferase reporter gene in transfected myometrial cell lines. Our investigation revealed two cis-acting regulatory elements within this fragment. Deletion of a region extending from −102 to −92 led to an increase of the promoter activity by greater than 10-fold, indicating a presence of a repressor element. Deletion of a region extending from −72 to −62 caused a decrease of the promoter activity of a similar extent, implying the existence of a positive element. Electrophoretic mobility shift assays demonstrated that synthetic oligonucleotides derived from these two small regions can each bind with a nuclear protein(s) prepared from myometrial cells, and an introduction of three and two base substitutions into each of these oligomers was sufficient to abolish their protein binding capability. These same mutations, when incorporated in the chloramphenicol acetyltransferase constructs, diminished regulatory functions of the negative and positive elements, and the protein(s) that bind to these functional elements was found in several tissues known to express cx43 gene.
大鼠妊娠期间参与胎母通讯的三种间隙连接基因产物的时空表达。
DOI：
10.1242/dev.113.1.165
发表时间：
1991
期刊：
Development (Cambridge, England)
影响因子：
--
作者：
Risek,B;Gilula,NB
通讯作者：
Gilula,NB
编码连接蛋白 43 的小鼠 Cx43 基因的结构、序列和表达。
影响因子：
3.5
作者：
Sullivan,R;Ruangvoravat,C;Joo,D;Morgan,J;Wang,BL;Wang,XK;Lo,CW
通讯作者：
Lo,CW
原代人平滑肌细胞的永生化。
DOI：
10.1073/pnas.89.4.1224
发表时间：
1992
影响因子：
11.1
作者：
Perez-Reyes,N;Halbert,CL;Smith,PP;Benditt,EP;McDougall,JK
通讯作者：
McDougall,JK