title: Second intron of mouse nestin gene directs its expression in pluripotent embryonic carcinoma cells through POU factor binding site.
abstract: 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 type-specific 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.
authors.affiliation: Laboratory of Stem Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
authors.forename: Zhi-Gang
authors.initials: ZG
authors.lastname: Jin
authors.forename: Li
authors.initials: L
authors.lastname: Liu
authors.forename: Hua
authors.initials: H
authors.lastname: Zhong
authors.forename: Ke-Jing
authors.initials: KJ
authors.lastname: Zhang
authors.forename: Yong-Feng
authors.initials: YF
authors.lastname: Chen
authors.forename: Wei
authors.initials: W
authors.lastname: Bian
authors.forename: Le-Ping
authors.initials: LP
authors.lastname: Cheng
authors.forename: Nai-He
authors.initials: NH
authors.lastname: Jing
mesh_terms: D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D002277:Carcinoma / Q000473:pathology; D004247:DNA / Q000235:genetics / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D007381:Intermediate Filament Proteins / Q000235:genetics* / Q000378:metabolism; D007438:Introns / Q000235:genetics*; D051379:Mice; D009154:Mutation; D009419:Nerve Tissue Proteins / Q000235:genetics* / Q000378:metabolism; D064231:Nestin; D050809:POU Domain Factors / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000378:metabolism*; D014407:Tumor Cells, Cultured
chemical_list: D007381:Intermediate Filament Proteins; D009419:Nerve Tissue Proteins; C577485:Nes protein, mouse; D064231:Nestin; D050809:POU Domain Factors; D004247:DNA