结合水分子参与雌激素受体和雌激素效应元件的体外结合  

Participation of Water in the Binding of Estrogen Receptor with Estrogen Responsive Element in vitro

作　　者：朱国璋[1] 唐国庆[1] 阮康成 龚岳亭 张永莲[1] 

机构地区：[1]中国科学院上海生物化学研究所分子生物学国家重点实验室

出　　处：《生物化学与生物物理学报》1998年第1期86-90,共5页

基　　金：国家自然科学基金

摘　　要：研究证明在许多生物大分子之间的相互作用中都有水分子的参与，但水分子是否参与真核基因转录调控中反式作用因子和顺式作用元件之间的结合尚未见报道。本文以雌激素受体（ＥＲ）同３２ｂｐ雌激素效应元件（ＥＲＥ）体外结合为模型，对结合水分子的参与进行探索性研究。在凝胶阻滞分析反应系统中加入不同浓度的渗透剂甘油、蔗糖、二甲基甲酰胺、二甲基亚砜，以调节反应系统的渗透压，随着渗透剂浓度的增大，ＥＲ－ＥＲＥ的结合也随着增加；相反，当给反应系统一个逐渐增大的静水压时，ＥＲ－ＥＲＥ的结合却逐渐减弱，撤除静水压后，ＥＲ－ＥＲＥ的结合随着时间的推移又逐渐恢复到原有的水平。结果提示结合水分子参与了ＥＲ－ＥＲＥ的结合，可能在真核基因转录调控中起一定的作用。Many reports have showed that bound water was involved in the interaction between/among the macromolecules. However, it has not been reported whether bound water is also involved in the binding of transfactors and ciselements in the regulation of the eukaryotic gene transcription or not. Preliminary studies have been made on the effect of bound water on the binding of estrogen receptor with estrogen responsive element in vitro. In the gel retardation assay using the cytosol extract of rat uterus as the supplier of estrogen receptor and 32 bp oligonucleotide containing a concensus vitellogenin A2 ERE as the probe, various cosolvents, such as glycerol, sucrose, Ndimethylformamide and dimethylsulfoxide, were added respectively to the reaction mixture in varying concentrations to regulate the osmotic pressure. The results indicated that the binding of ERERE was enhanced with the increase in the final concentration of these individual cosolvents. On the other hand, when the reaction was carried out under an increasing hydrostatic pressure, the ERERE binding was decreased sharply. After decompression the binding of ERERE was gradually restored to the normal level with the lapse of time. These results suggested that bound water was directly involved in the binding of ERERE and may play an important role in the regulation of the eukaryotic gene transcription.

关 键 词：结合水分子 雌激素 受体 效应元件 渗透压 

分 类 号：Q579.13[生物学—生物化学]