title: PRDM14 promotes active DNA demethylation through the ten-eleven translocation (TET)-mediated base excision repair pathway in embryonic stem cells.
abstract: Ten-eleven translocation (TET) proteins oxidize 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC) and 5-carboxylcytosine (5caC). 5fC and 5caC can be excised and repaired by the base excision repair (BER) pathway, implicating 5mC oxidation in active DNA demethylation. Genome-wide DNA methylation is erased in the transition from metastable states to the ground state of embryonic stem cells (ESCs) and in migrating primordial germ cells (PGCs), although some resistant regions become demethylated only in gonadal PGCs. Understanding the mechanisms underlying global hypomethylation in naive ESCs and developing PGCs will be useful for realizing cellular pluripotency and totipotency. In this study, we found that PRDM14, the PR domain-containing transcriptional regulator, accelerates the TET-BER cycle, resulting in the promotion of active DNA demethylation in ESCs. Induction of Prdm14 expression transiently elevated 5hmC, followed by the reduction of 5mC at pluripotency-associated genes, germline-specific genes and imprinted loci, but not across the entire genome, which resembles the second wave of DNA demethylation observed in gonadal PGCs. PRDM14 physically interacts with TET1 and TET2 and enhances the recruitment of TET1 and TET2 at target loci. Knockdown of TET1 and TET2 impaired transcriptional regulation and DNA demethylation by PRDM14. The repression of the BER pathway by administration of pharmacological inhibitors of APE1 and PARP1 and the knockdown of thymine DNA glycosylase (TDG) also impaired DNA demethylation by PRDM14. Furthermore, DNA demethylation induced by PRDM14 takes place normally in the presence of aphidicolin, which is an inhibitor of G1/S progression. Together, our analysis provides mechanistic insight into DNA demethylation in naive pluripotent stem cells and developing PGCs.
authors: Department of Bioscience, School of Science and Technology, Kwansei Gakuin University, 2-1 Gakuen, Sanda, Hyogo 669-1337, Japan.
authors: Naoki
authors: N
authors: Okashita
authors: Yuichi
authors: Y
authors: Kumaki
authors: Kuniaki
authors: K
authors: Ebi
authors: Miyuki
authors: M
authors: Nishi
authors: Yoshinori
authors: Y
authors: Okamoto
authors: Megumi
authors: M
authors: Nakayama
authors: Shota
authors: S
authors: Hashimoto
authors: Tomohumi
authors: T
authors: Nakamura
authors: Kaoru
authors: K
authors: Sugasawa
authors: Nakao
authors: N
authors: Kojima
authors: Tatsuyuki
authors: T
authors: Takada
authors: Masaki
authors: M
authors: Okano
authors: Yoshiyuki
authors: Y
authors: Seki
mesh_terms: D000818:Animals; D019175:DNA Methylation / Q000235:genetics / Q000502:physiology; D004260:DNA Repair / Q000235:genetics / Q000502:physiology*; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D049308:Dioxygenases; D053595:Embryonic Stem Cells / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D018392:Genomic Imprinting; D005854:Germ Cells / Q000378:metabolism; D051379:Mice; D039904:Pluripotent Stem Cells / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D016601:RNA-Binding Proteins; D015398:Signal Transduction; D044502:Thymine DNA Glycosylase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: D004268:DNA-Binding Proteins; C531066:Prdm14 protein, mouse; D011518:Proto-Oncogene Proteins; D016601:RNA-Binding Proteins; C540839:TET1 protein, mouse; D014157:Transcription Factors; D049308:Dioxygenases; C551914:Tet2 protein, mouse; D044502:Thymine DNA Glycosylase
keywords: Base excision repair (BER); DNA demethylation; Embryonic stem cells; Mouse; Ten-eleven translocation (TET)