1.SUMO-2/3 can covalently bind to progesterone receptor B to regulate its transcriptional activity.
Bai-yu HAN ; Fa-ceng LI ; Long CHENG ; Xiao-jie XU ; Kai JIANG ; Jie FU ; Yong-jian HAN ; Zhao-hui LV ; Jing-tao DOU ; Hao ZHANG ; Qi-nong YE
Journal of Southern Medical University 2011;31(9):1493-1497
OBJECTIVETo investigate whether progesterone receptor B (PRB) can be sumoylated by SUMO-2/3 and the effect of sumoylation on PRB transcriptional activity.
METHODSSUMO-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.
RESULTSpcDNA3FLAG-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.
CONCLUSIONPRB can be sumoylated by SUMO-2/3 and its function is regulated by this modification.
Animals ; Cell Line ; Humans ; Plasmids ; genetics ; Receptors, Progesterone ; genetics ; metabolism ; Small Ubiquitin-Related Modifier Proteins ; genetics ; metabolism ; Transcription, Genetic ; Transfection ; Ubiquitination ; Ubiquitins ; genetics ; metabolism
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