HDAC1对ERα和ERβ蛋白水平的调控  

Modulation of ERα and ERβ protein levels by HDAC1

作　　者：常庆[1] 韩聚强[1] 王晓辉[1] 姜艳超[1] 丁丽华[1] 李杰之[1] 陈宗德[2] 李熔[1] 叶棋浓[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100850 [2]郑州大学基础医学院,郑州450052

出　　处：《军事医学科学院院刊》2006年第4期329-332,共4页Bulletin of the Academy of Military Medical Sciences

基　　金：国家自然科学基金(30530320;30428012和30470378)

摘　　要：目的:构建带编码HA标签的hdac1基因真核表达载体,研究组蛋白去乙酰化酶1(HDAC1)对雌激素受体α和β(ERα和ERβ)蛋白水平的影响。方法:用提取的RNA产物进行逆转录合成cDNA,利用PCR技术扩增出hdac1基因完整编码区序列,通过DNA重组技术构建含编码hdac1DNA片段的真核表达载体pcDNA3-HA-hdac1;利用GST沉降(pu ll-down)实验观察HDAC1是否能特异结合ERα或ERβ;又将pcDNA3-HA-hdac1重组质粒分别和ERα表达载体或ERβ表达载体质粒共转染293T细胞,观察HDAC1对ERα和ERβ蛋白水平的影响。结果:转染pcDNA3-HA-hdac1重组质粒的哺乳动物细胞表达了与预期相对分子质量大小一致的HDAC1蛋白;GST沉降实验表明,HDAC1蛋白与ERα和ERβ均结合;共转染实验观察到HDAC1可使ERα蛋白水平降低,而对ERβ没有影响。结论:HDAC1与ERα和ERβ均结合,但其对ER s的作用具有亚型特异性,ERα和ERβ蛋白水平可能受不同类型的组蛋白去乙酰化酶(HDACs)的调控。Objectives： To construct the hdacleukaryotic expression vector with HA tag, and to investigate the effect of histone deacetylases 1 （HDAC1） on ERα and ERβ protein levels. Methods：The PCR amplified DNA fragment containing entire hdacl coding region was cloned into the pcDNA3-HA vector, resulting in the pcDNA3-HA-hdac1 recombinant plasmid. The interaction between HDAC1 and ERs（ ERα and ERβ） was tested by using GST pull-down assay. 293T Cells were co-transfected with HA-tagged hdac1 recombinant plasmid and ERα or ERβ expression plasmid to observe the change in ERα and ERβ protein levels. Results：Western blot analysis showed that 293T cells transfected with pcDNA3-HA-hdac1 expressed HDAC1 protein as expected. GST pull-down assay suggested that HDAC1 interact with both ERα and ERβ. Cotransfection experiments indicated that HDAC1 reduced the protein level of ERα, but not ERβ. Conclusion： This study suggested that the effect of HDAC1 on ER protein level be ER subtype-specific, though HDAC1 could interact with both ERα and ERβ. The protein levels of ERα and ERβ may be modulated by different kinds of HDACs.

关 键 词：HDAC1 受体 雌激素 基因表达 蛋白水平 

分 类 号：Q75[生物学—分子生物学]