title: DNA methylation is required for silencing of ant4, an adenine nucleotide translocase selectively expressed in mouse embryonic stem cells and germ cells.
abstract: The capacity for cellular differentiation is governed not only by the repertoire of available transcription factors but by the accessibility of cis-regulatory elements. Studying changes in epigenetic modifications during stem cell differentiation will help us understand how cells maintain or lose differentiation potential. We investigated changes in DNA methylation during the transition of pluripotent embryonic stem cells (ESCs) into differentiated cell types. Using a methylation-sensitive restriction fingerprinting method, we identified a novel adenine nucleotide (ADP/ATP) translocase gene, Ant4, that was selectively hypomethylated and expressed in undifferentiated mouse ESCs. In contrast to other pluripotent stem cell-specific genes such as Oct-4 and Nanog, the Ant4 gene was readily derepressed in differentiated cells after 5-aza-2'-deoxycytidine treatment. Moreover, expression of de novo DNA methyltransferases Dnmt3a and Dnmt3b was essential for repression and DNA methylation of the Ant4 gene during ESC differentiation. Although the deduced amino acid sequence of Ant4 is highly homologous to the previously identified Ant isoforms, the expression of Ant4 was uniquely restricted to developing gametes in adult mice, and its promoter hypomethylation was observed only in testis. Additionally, Ant4 was expressed in primordial germ cells. These data indicate that Ant4 is a pluripotent stem cell- and germ cell-specific isoform of adenine nucleotide translocase in mouse and that DNA methylation plays a primary role in its transcriptional silencing in somatic cells.
authors.affiliation: Department of Pathology, University of Florida College of Medicine, 1600 SW Archer Road, Gainesville, Florida 32610, USA.
authors.forename: Nemanja
authors.initials: N
authors.lastname: Rodić
authors.forename: Masahiro
authors.initials: M
authors.lastname: Oka
authors.forename: Takashi
authors.initials: T
authors.lastname: Hamazaki
authors.forename: Matthew R
authors.initials: MR
authors.lastname: Murawski
authors.forename: Marda
authors.initials: M
authors.lastname: Jorgensen
authors.forename: Danielle M
authors.initials: DM
authors.lastname: Maatouk
authors.forename: James L
authors.initials: JL
authors.lastname: Resnick
authors.forename: En
authors.initials: E
authors.lastname: Li
authors.forename: Naohiro
authors.initials: N
authors.lastname: Terada
mesh_terms: D000818:Animals; D001483:Base Sequence; D002454:Cell Differentiation / Q000235:genetics; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000172:deficiency / Q000235:genetics; D019175:DNA Methylation; D000090205:DNA Methyltransferase 3A; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism; D005786:Gene Expression Regulation; D020868:Gene Silencing; D008297:Male; D051379:Mice; D000226:Mitochondrial ADP, ATP Translocases / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D013094:Spermatozoa / Q000201:enzymology; D013234:Stem Cells / Q000166:cytology / Q000201:enzymology*; D013737:Testis / Q000166:cytology; D000097584:DNA Methyltransferase 3B
chemical_list: D004248:DNA (Cytosine-5-)-Methyltransferases; D000090205:DNA Methyltransferase 3A; D000226:Mitochondrial ADP, ATP Translocases; D012333:RNA, Messenger; D000097584:DNA Methyltransferase 3B; C000715535:Dnmt3a protein, mouse