title: Oct4 and the small molecule inhibitor, SC1, regulates Tet2 expression in mouse embryonic stem cells.
abstract: The ten eleven translocation (Tet) family of proteins includes three members (Tet1-3), all of which have the capacity to convert 5-methylcytosine to 5-hydroxymethylcytosine in a 2-oxoglutarate- and Fe(II)-dependent manner. Tet1 and Tet2 are highly expressed in undifferentiated embryonic stem cells (ESCs), and this expression decreases upon differentiation. Notably, the expression patterns of Tet1 and Tet2 in ESCs parallels that of pluripotency genes. To date, however, the mechanisms underlying the regulation of Tet gene expression in ESCs remain largely unexplored. Here we report that the pluripotency transcription factor, Oct4, directly regulates the expression of Tet2. Using RNAi, real time quantitative PCR, dual-luciferase reporter assays and electrophoretic mobility shift assays, we show that Oct4 promotes Tet2 transcription by binding to consensus sites in the proximal promoter region. Furthermore, we explored the role of the small molecule inhibitor, SC1 (pluripotin) on Tet gene expression. We show that SC1 promotes Tet3 expression, but represses Tet1 and Tet2 expression. Our findings indicate that Tet2 are crucial downstream targets of the pluripotency factor Oct4, and highlight a role for Oct4 in the regulation of DNA methylation in ESCs. In addition, these findings also provide a new insight into drug-mediated gene regulation.
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, Key Laboratory of Animal Biotechnology, Ministry of Agriculture, 3 Taicheng Road, Yangling 712100, China.
authors.forename: Yongyan
authors.initials: Y
authors.lastname: Wu
authors.forename: Zekun
authors.initials: Z
authors.lastname: Guo
authors.forename: Ye
authors.initials: Y
authors.lastname: Liu
authors.forename: Bo
authors.initials: B
authors.lastname: Tang
authors.forename: Yi
authors.initials: Y
authors.lastname: Wang
authors.forename: Liping
authors.initials: L
authors.lastname: Yang
authors.forename: Juan
authors.initials: J
authors.lastname: Du
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D044503:5-Methylcytosine / Q000378:metabolism / Q000031:analogs & derivatives; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D003596:Cytosine / Q000031:analogs & derivatives / Q000378:metabolism; D019175:DNA Methylation / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D049308:Dioxygenases; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D051379:Mice; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D011720:Pyrazoles / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology
chemical_list: D004268:DNA-Binding Proteins; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D011518:Proto-Oncogene Proteins; D011720:Pyrazoles; D011743:Pyrimidines; C515825:SC1 compound; C540839:TET1 protein, mouse; C011865:5-hydroxymethylcytosine; D044503:5-Methylcytosine; D003596:Cytosine; D049308:Dioxygenases; C551914:Tet2 protein, mouse; C551915:Tet3 protein, mouse