[小泛素样修饰蛋白2/3可与孕激素受体B共价结合以调节其转录活性]

[SUMO-2/3 can covalently bind to progesterone receptor B to regulate its transcriptional activity].

作者信息

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

机构信息

Department of Endocrinology, General Hospital of PLA, Beijing 100853, China.

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2011 Sep;31(9):1493-7.

PMID:21945751

Abstract

OBJECTIVE

To investigate whether progesterone receptor B (PRB) can be sumoylated by SUMO-2/3 and the effect of sumoylation on PRB transcriptional activity.

METHODS

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.

RESULTS

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.

CONCLUSION

PRB can be sumoylated by SUMO-2/3 and its function is regulated by this modification.

摘要

目的

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。

方法

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。