Han Bai-yu, Li Fa-ceng, Cheng Long, Xu Xiao-jie, Jiang Kai, Fu Jie, Han Yong-jian, Lv Zhao-hui, Dou Jing-tao, Zhang Hao, Ye Qi-nong
Department of Endocrinology, General Hospital of PLA, Beijing 100853, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2011 Sep;31(9):1493-7.
To investigate whether progesterone receptor B (PRB) can be sumoylated by SUMO-2/3 and the effect of sumoylation on PRB transcriptional activity.
SUMO-2/3 cDNA was amplified from MCF-7 cDNA and cloned into the eukaryotic expression vector pcDNA3-FLAG. The plasmid pXJ40-myc-PRB was cotransfected with pcDNA3FLAG-SUMO2, pcDNA3FLAG-SUMO3 or the mock control into 293T cells, and PRB sumoylation was detected by immunoprecipitation and Western blotting. The effect of PRB sumoylation on its transcriptional activity was determined using reporter luciferase assay.
pcDNA3FLAG-SUMO2 and pcDNA3FLAG-SUMO3 vectors were successfully constructed. SUMO-2/3 could bind covalently to PRB and increase its transcriptional dependent on the presence of progesterone.
PRB can be sumoylated by SUMO-2/3 and its function is regulated by this modification.
研究孕激素受体B（PRB）是否可被SUMO-2/3 进行类泛素化修饰以及这种修饰对PRB转录活性的影响。
从MCF-7 cDNA中扩增SUMO-2/3 cDNA，并克隆到真核表达载体pcDNA3-FLAG中。将质粒pXJ40-myc-PRB与pcDNA3FLAG-SUMO2、pcDNA3FLAG-SUMO3或空载体对照共转染至293T细胞中，通过免疫沉淀和蛋白质印迹法检测PRB的类泛素化修饰情况。使用荧光素酶报告基因检测法确定PRB类泛素化修饰对其转录活性的影响。
成功构建了pcDNA3FLAG-SUMO2和pcDNA3FLAG-SUMO3载体。SUMO-2/3能够与PRB共价结合，并在孕激素存在的情况下增强其转录活性。
PRB可被SUMO-2/3进行类泛素化修饰，且其功能受这种修饰的调控。
