title: Dppa3 expression is critical for generation of fully reprogrammed iPS cells and maintenance of Dlk1-Dio3 imprinting.
abstract: Reprogramming of mouse somatic cells into induced pluripotent stem cells (iPSCs) often generates partially reprogrammed iPSCs (pre-iPSCs), low-grade chimera forming iPSCs (lg-iPSCs) and fully reprogrammed, high-grade chimera production competent iPSCs (hg-iPSCs). Lg-iPSC transcriptome analysis revealed misregulated Dlk1-Dio3 cluster gene expression and subsequently the imprinting defect at the Dlk1-Dio3 locus. Here, we show that germ-cell marker Dppa3 is present only in lg-iPSCs and hg-iPSCs, and that induction with exogenous Dppa3 enhances reprogramming kinetics, generating all hg-iPSCs, similar to vitamin C (Vc). Conversely, Dppa3-null fibroblasts show reprogramming block at pre-iPSCs state and Dlk1-Dio3 imprinting defect. At the molecular level, we show that Dppa3 is associated with Dlk1-Dio3 locus and identify that Dppa3 maintains imprinting by antagonizing Dnmt3a binding. Our results further show molecular parallels between Dppa3 and Vc in Dlk1-Dio3 imprinting maintenance and suggest that early activation of Dppa3 is one of the cascades through which Vc facilitates the generation of fully reprogrammed iPSCs.
authors.affiliation: Institute of Human Genetics, University of Goettingen, Heinrich-Dueker-Weg 12, 37073 Goettingen, Germany.
authors.forename: Xingbo
authors.initials: X
authors.lastname: Xu
authors.affiliation: Institute of Human Genetics, University of Goettingen, Heinrich-Dueker-Weg 12, 37073 Goettingen, Germany.
authors.forename: Lukasz
authors.initials: L
authors.lastname: Smorag
authors.affiliation: Department of Bio-Science, Nagahama Institute of Bio-Science and Technology, Shiga 526-0829, Japan.
authors.forename: Toshinobu
authors.initials: T
authors.lastname: Nakamura
authors.affiliation: Department of Biosciences, Kitasato University School of Science, Kanagawa 252-0373, Japan.
authors.forename: Tohru
authors.initials: T
authors.lastname: Kimura
authors.affiliation: Department of Cellular and Molecular Immunology, University of Goettingen, Humboldtallee 34, 37073 Goettingen, Germany.
authors.forename: Ralf
authors.initials: R
authors.lastname: Dressel
authors.affiliation: Institute of Human Genetics, Johannes Gutenberg-University Mainz, Langenbeckstraße 1, 55131 Mainz, Germany.
authors.forename: Antje
authors.initials: A
authors.lastname: Fitzner
authors.affiliation: Institute of Human Genetics, University of Goettingen, Heinrich-Dueker-Weg 12, 37073 Goettingen, Germany.
authors.forename: Xiaoying
authors.initials: X
authors.lastname: Tan
authors.affiliation: Institute of Human Genetics, Johannes Gutenberg-University Mainz, Langenbeckstraße 1, 55131 Mainz, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Linke
authors.affiliation: Institute of Human Genetics, Johannes Gutenberg-University Mainz, Langenbeckstraße 1, 55131 Mainz, Germany.
authors.forename: Ulrich
authors.initials: U
authors.lastname: Zechner
authors.affiliation: Institute of Human Genetics, University of Goettingen, Heinrich-Dueker-Weg 12, 37073 Goettingen, Germany.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Engel
authors.affiliation: Institute of Human Genetics, University of Goettingen, Heinrich-Dueker-Weg 12, 37073 Goettingen, Germany.
authors.forename: D V Krishna
authors.initials: DV
authors.lastname: Pantakani
mesh_terms: D000818:Animals; D001205:Ascorbic Acid / Q000378:metabolism; D002135:Calcium-Binding Proteins; D002868:Chromosomal Proteins, Non-Histone; D003433:Crosses, Genetic; D019175:DNA Methylation; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D018392:Genomic Imprinting; D005854:Germ Cells / Q000166:cytology; D049452:Green Fluorescent Proteins / Q000378:metabolism; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism*; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism*; D007453:Iodide Peroxidase / Q000378:metabolism*; D007700:Kinetics; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008822:Mice, Transgenic; D011485:Protein Binding; D012097:Repressor Proteins / Q000378:metabolism*; D012190:Retroviridae / Q000378:metabolism
chemical_list: D002135:Calcium-Binding Proteins; D002868:Chromosomal Proteins, Non-Histone; C512925:Dlk1 protein, mouse; C462180:Dppa3 protein, mouse; D036341:Intercellular Signaling Peptides and Proteins; D012097:Repressor Proteins; D049452:Green Fluorescent Proteins; C117274:iodothyronine deiodinase type III; D007453:Iodide Peroxidase; D001205:Ascorbic Acid