Second Intron of Mouse Nestin Gene Directs its Expression in Pluripotent Embryonic Carcinoma Cells through POU Factor Binding Site  

作　　者：Zhi-Gang JIN Li LIU Hua ZHONG Ke-Jing ZHANG Yong-Feng CHEN Wei BIAN Le-Ping CHENG Nai-He JING 

机构地区：[1]Laboratory of Stem Cell Biology, Laboratory of Molecular Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China

出　　处：《Acta Biochimica et Biophysica Sinica》2006年第3期207-212,共6页生物化学与生物物理学报（英文版）

基　　金：the National Natural Science Foundation of China （No. 90208011, 30300174, 30470856 , 30421005）; the National Key Basic Research and Development Program of China （No. 2002CB713802 , 2005CB522703） ; the Key Project of Basic Science Research of Shanghai （No. 04DZ14005 , 04DZ05608）.

摘　　要：Nestin, an intermediate filament protein, is expressed in the neural stem cells of the developing central nervous system. This tissue-specific expression is driven by the neural stem cell-specific enhancer in the second intron of the nestin gene. In this study, we showed that the mouse nestin gene was expressed in pluripotent embryonic carcinoma （EC） P19 and F9 cells, not in the differentiated cell types. This cell typespecific expression was conferred by the enhancer in the second intron. Mutation of the conserved POU factor-binding site in the enhancer abolished the reporter gene expression in EC cells. Oct4, a Class V POU factor, was found to be coexpressed with nestin in EC cells. Electrophoretic mobility-shift assays and supershift assays showed that a unique protein-DNA complex was formed specifically with nuclear extracts of EC cells, and Oct4 protein was included. Together, these results suggest the functional relevance between the conserved POU factor-binding site and the expression of the nestin gene in pluripotent EC cells.Nestin, an intermediate filament protein, is expressed in the neural stem cells of the developing central nervous system. This tissue-specific expression is driven by the neural stem cell-specific enhancer in the second intron of the nestin gene. In this study, we showed that the mouse nestin gene was expressed in pluripotent embryonic carcinoma （EC） P19 and F9 cells, not in the differentiated cell types. This cell typespecific expression was conferred by the enhancer in the second intron. Mutation of the conserved POU factor-binding site in the enhancer abolished the reporter gene expression in EC cells. Oct4, a Class V POU factor, was found to be coexpressed with nestin in EC cells. Electrophoretic mobility-shift assays and supershift assays showed that a unique protein-DNA complex was formed specifically with nuclear extracts of EC cells, and Oct4 protein was included. Together, these results suggest the functional relevance between the conserved POU factor-binding site and the expression of the nestin gene in pluripotent EC cells.

关 键 词：nestin embryonic carcinoma cells ENHANCER POU factor 

分 类 号：R73[医药卫生—肿瘤]