期刊
BIOCHIMICA ET BIOPHYSICA ACTA-GENE REGULATORY MECHANISMS
卷 1799, 期 8, 页码 546-554出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.bbagrm.2010.07.003
关键词
Estrogen response element; Estrogen receptor; Vitellogenin; Rainbow trout; Transfection assay; Fluorescence anisotropy
资金
- Le Ministere de l'Enseignement Superieur et de la Recherche
- Le Comite du Morbihan de la Ligue Nationale Contre le Cancer
In most of oviparous animals, vitellogenins (VTG) are the major egg yolk precursors. They are produced in the liver under the control of estrogens. In rainbow trout (Oncorhynchus mykiss). the vtg genes cluster contains an unusually large number of almost identical gene copies. In order to identify the regulatory elements in their promoters, we used a combination of reporter plasmids containing genomic sequences including putative estrogen response elements (EREs) and we performed transient transfection assays in MCF-7 and yeast cells. We found a functional ERE corresponding to the sequence GGGGCAnnnTAACCT (rtvtgERE), which differs from the consensus ERE (ERE) by three base pairs. This non-palindromic ERE is located in the env gene of a retrotransposon relic, 180 base pairs upstream of the transcriptional start site. Fluorescence anisotropy experiments confirmed that the purified human estrogen receptor alpha (hER alpha) can specifically bind to rtvtgERE. Furthermore, we observe that the stability of hER alpha-EREcs and hER alpha-rtvtgERE complexes is similar with equilibrium dissociation constants of 3.0 nM and 6.2 nM respectively, under our experimental conditions. Additionally, this rtvtgERE sequence displays a high E2-responsiveness through ER activation in cellulo. In the rainbow trout, the functional ERE (rtvtgERE) lies within promoter sequences which are mostly composed of sequences derived from transposable elements (TEs), which therefore may have acted as an evolutionary buffer to secure the proper expression of these genes. (C) 2010 Elsevier B.V. All rights reserved.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据