An ESRG-interacting protein, COXII, is involved in pro-apoptosis of human embryonic stem cells.
Human embryonic stem cells(hESC) posses very promising application perspective in clinical transplant therapies for their characteristics of self-renewal and pluripotency. So efforts focusing on the mechanisms of the two characteristics are extremely important. ESRG, first identified by our group, is a candidate stemness gene of hESC for its much higher expression level in hESC comparing to that in 7-day embryoid bodies(EBs). Here, the proteins interacted with ESRG and its functions in hESC were explored. Yeast two-hybrid (Y2H) screening system was adopted to explore the interacting proteins of ESRG. Then Co-IP was performed to confirm the interactions between candidate proteins and ESRG. At last, the functions of validated interacting protein were explored by RNA interference(RNAi) and Western blot(WB). There were no autonomous activation and toxicity in the Y2H system, which verified its availability. Four candidate proteins, AAMP, DDT, GNB2L1 and COXII, were discovered, and the interaction between ESRG and COXII was ultimately confirmed. The expression of COXII in hESC was suppressed by siRNA, and the inhibited mitochondrial apoptosis was observed in hESC with downregulated COXII expression. Our work first validated the interaction between ESRG and COXII, and demonstrated that COXII serves as a pro-apoptotic protein in hESC. The results implied that ESRG may play an important role in regulating the apoptosis of hESC by interacting with COXII, and thus contribute a lot to the maintenance of hESC characteristics.
Cancer Research Institute, Collaborative Innovation Center for Cancer Medicine, Key Laboratory for Carcinogenesis of Chinese Ministry of Health, School of Basic Medical Sciences, Central South University, Xiangya Road 110, 410078 Changsha, Hunan, PR China.
Jia
J
Shi
Cancer Research Institute, Collaborative Innovation Center for Cancer Medicine, Key Laboratory for Carcinogenesis of Chinese Ministry of Health, School of Basic Medical Sciences, Central South University, Xiangya Road 110, 410078 Changsha, Hunan, PR China. Electronic address: rencaiping@csu.edu.cn.
Caiping
C
Ren
Cancer Research Institute, Collaborative Innovation Center for Cancer Medicine, Key Laboratory for Carcinogenesis of Chinese Ministry of Health, School of Basic Medical Sciences, Central South University, Xiangya Road 110, 410078 Changsha, Hunan, PR China.
Hui
H
Liu
Cancer Research Institute, Collaborative Innovation Center for Cancer Medicine, Key Laboratory for Carcinogenesis of Chinese Ministry of Health, School of Basic Medical Sciences, Central South University, Xiangya Road 110, 410078 Changsha, Hunan, PR China.
Lei
L
Wang
Cancer Research Institute, Collaborative Innovation Center for Cancer Medicine, Key Laboratory for Carcinogenesis of Chinese Ministry of Health, School of Basic Medical Sciences, Central South University, Xiangya Road 110, 410078 Changsha, Hunan, PR China.
Bin
B
Zhu
Cancer Research Institute, Collaborative Innovation Center for Cancer Medicine, Key Laboratory for Carcinogenesis of Chinese Ministry of Health, School of Basic Medical Sciences, Central South University, Xiangya Road 110, 410078 Changsha, Hunan, PR China.
Wei
W
Huang
Cancer Research Institute, Collaborative Innovation Center for Cancer Medicine, Key Laboratory for Carcinogenesis of Chinese Ministry of Health, School of Basic Medical Sciences, Central South University, Xiangya Road 110, 410078 Changsha, Hunan, PR China.
Weidong
W
Liu
Cancer Research Institute, Collaborative Innovation Center for Cancer Medicine, Key Laboratory for Carcinogenesis of Chinese Ministry of Health, School of Basic Medical Sciences, Central South University, Xiangya Road 110, 410078 Changsha, Hunan, PR China.
Jie
J
Liu
Cancer Research Institute, Collaborative Innovation Center for Cancer Medicine, Key Laboratory for Carcinogenesis of Chinese Ministry of Health, School of Basic Medical Sciences, Central South University, Xiangya Road 110, 410078 Changsha, Hunan, PR China.
Yanyu
Y
Liu
Department of Gynaecology and Obstetrics, The Second Xiangya Hospital, Central South University, 139 Middle Renmin Road, Changsha, Hunan 410011, PR China.
Xiaomeng
X
Xia
Cancer Research Institute, Collaborative Innovation Center for Cancer Medicine, Key Laboratory for Carcinogenesis of Chinese Ministry of Health, School of Basic Medical Sciences, Central South University, Xiangya Road 110, 410078 Changsha, Hunan, PR China.
Rong
R
Xu
Department of Neurosurgery, Xiangya Hospital, Central South University, 87 Xiangya Road, Changsha, Hunan 410008, China. Electronic address: jiangxingjun@sina.com.
Xingjun
X
Jiang
D017209:Apoptosis; D001483:Base Sequence; D017931:DNA Primers; D003576:Electron Transport Complex IV / Q000378:metabolism*; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000201:enzymology; D006801:Humans; D016133:Polymerase Chain Reaction
D017931:DNA Primers; D003576:Electron Transport Complex IV
COXII; ESRG; Mitochondrial apoptosis; hESC