ETO Family Protein Mtgr1 Mediates Prdm14 Functions in Stem Cell Maintenance
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE74547 | GEO | 2016/01/01
SECONDARY ACCESSION(S): PRJNA300681
REPOSITORIES: GEO
Similar Datasets
Project description:Prdm14 is a sequence-specific transcriptional regulator of embryonic stem cell (ESC) pluripotency and primordial germ cell (PGC) formation. It exerts its function, at least in part, through repressing genes associated with epigenetic modification and cell differentiation. Here, we show that this repressive function is mediated through an ETO-family co-repressor Mtgr1, which tightly binds to the pre-SET/SET domains of Prdm14 and co-occupies its genomic targets in mouse ESCs. Structure-guided point mutants abrogated the Prdm14-Mtgr1 association and disrupted Prdm14's function in mESC gene expression and PGC formation in vitro. Altogether, our work uncovers the molecular mechanism underlying Prdm14-mediated repression. Examination of Prdm14 and Mtgr1 occupancy by ChIP-seq and effects on gene expression in mouse embryonic stem cells
2016-07-03 | E-GEOD-74547 | biostudies-arrayexpress
Project description:Primordial germ cells (PGCs), the founder cells of the germline, are specified in pre-gastrulating embryos in mammals, and subsequently migrate towards gonads to mature into functional gametes. Here, we investigated PGC development in rats, by genetically modifying Prdm14, a unique marker and a critical PGC transcriptional regulator. We trace PGC development in rats, for the first time, from specification until sex determination stage in fetal gonads using Prdm14 H2BVenus knock-in rats. We uncover that Prdm14’s crucial role in PGC specification is conserved between rat and mice, by analyzing Prdm14 deficient rat embryos. Notably, loss of Prdm14 completely abrogates the PGC program: failure in maintenance and/or activation of germ cell markers and pluripotency genes. Finally, we profile the transcriptome of the postimplantation epiblast and all PGC stages in rat, to reveal enrichment of distinct gene sets at each transition point, thereby providing an accurate transcriptional time-line for rat PGC development. Thus, the novel genetically modified rats and data sets obtained in this study will advance our knowledge on conserved vs species-specific features for germline development in mammals.
2019-12-26 | GSE135388 | GEO
Project description:Prdm14 is a PR-domain and zinc-finger protein whose expression is restricted to the pluripotent cells of an early embryo, embryonic stem cells (ESCs), and germ cells. Here we show that Prdm14 safeguards mouse ESC maintenance by preventing induction of extraembryonic endoderm (ExEn) fates. Conversely, Prdm14 overexpression impairs ExEn differentiation during embryoid body (EB) formation. Prdm14 occupies and represses genomic loci encoding ExEn differentiation factors, while also binding to and promoting expression of genes associated with ESC self-renewal. Prdm14-bound genomic regions significantly overlap those occupied by Nanog and Oct4, are enriched in a chromatin signature associated with distal regulatory elements, and contain a unique DNA-sequence motif recognized by Prdm14 in vitro. Our work identifies Prdm14 as a new member of mouse ESC (mESC) transcriptional network, which plays a dual role as a context-dependent transcriptional repressor or activator at distal silencers and enhancers. [ChIP-seq] Genome-wide mapping of Prdm14 binding sites in mouse embryonic stem cells: A FLAG-HA tagged Prdm14 (FH-Prdm14) mESC line was established. FLAG-HA double ChIP (ChIP with FLAG antibody followed by ChIP with HA antibody) was performed with FH-Prdm14 mESCs (Prdm14-ChIPseq) and as a negative control, wildtype mESCs (FLAG-HA_ChIPseq). H3K4me1 ChIPseq in mouse ES cells. Using published H3K4me1 data, we found there is a correlation between Prdm14 binding and H3K4me1 marks. So we obtained our own H3K4me1 data, using the wildtype mESCs. [RNA-seq] Global RNAseq analysis of Prdm14 knockdown in mouse embryonic stem cells: Analysis of poly(A)+ RNA from mESCs treated with non-targeting control siRNA and Prdm14 siRNA.
2010-12-23 | E-GEOD-25409 | biostudies-arrayexpress
Project description:Prdm14 is a PR-domain and zinc-finger protein whose expression is restricted to the pluripotent cells of an early embryo, embryonic stem cells (ESCs), and germ cells. Here we show that Prdm14 safeguards mouse ESC maintenance by preventing induction of extraembryonic endoderm (ExEn) fates. Conversely, Prdm14 overexpression impairs ExEn differentiation during embryoid body (EB) formation. Prdm14 occupies and represses genomic loci encoding ExEn differentiation factors, while also binding to and promoting expression of genes associated with ESC self-renewal. Prdm14-bound genomic regions significantly overlap those occupied by Nanog and Oct4, are enriched in a chromatin signature associated with distal regulatory elements, and contain a unique DNA-sequence motif recognized by Prdm14 in vitro. Our work identifies Prdm14 as a new member of mouse ESC (mESC) transcriptional network, which plays a dual role as a context-dependent transcriptional repressor or activator at distal silencers and enhancers.
2010-12-23 | GSE25409 | GEO
Project description:PRDM14 belongs to the PR (PRDI-BF1 and RIZ) domain proteins (PRDM) family which is a subclass of the SET domain proteins, a common domain found in histone modifying enzymes. PRDM14 has been previously implicated to regulate self-renewal of hESCs as knock-down of PRDM14 induced expression of differentiation marker genes and altered the cellular morphology. We showed that PRDM14 directly regulates the expression of key pluripotency gene POU5F1. Genome-wide location profiling experiments revealed that PRDM14 co-localized extensively with other key transcription factors such as OCT4, NANOG and SOX2. More importantly, in a gain-of-function assay, we showed that PRDM14 is able to enhance the efficiency of reprogramming of human fibroblasts in conjunction with OCT4, SOX2 and KLF4. Hence, PRDM14 exemplifies a key transcription factor that is required for the maintenance of human ESC identity and the reacquisition of pluripotency in human somatic cells.
2010-10-20 | GSE22767 | GEO
Project description:PRDM14 belongs to the PR (PRDI-BF1 and RIZ) domain proteins (PRDM) family which is a subclass of the SET domain proteins, a common domain found in histone modifying enzymes. PRDM14 has been previously implicated to regulate self-renewal of hESCs as knock-down of PRDM14 induced expression of differentiation marker genes and altered the cellular morphology. We showed that PRDM14 directly regulates the expression of key pluripotency gene POU5F1. Genome-wide location profiling experiments revealed that PRDM14 co-localized extensively with other key transcription factors such as OCT4, NANOG and SOX2. More importantly, in a gain-of-function assay, we showed that PRDM14 is able to enhance the efficiency of reprogramming of human fibroblasts in conjunction with OCT4, SOX2 and KLF4. Hence, PRDM14 exemplifies a key transcription factor that is required for the maintenance of human ESC identity and the reacquisition of pluripotency in human somatic cells. ChIP-seq of PRDM14 in human ESCs
2010-10-20 | E-GEOD-22767 | biostudies-arrayexpress
Project description:PRDM14 is a crucial regulator of mouse primordial germ cells (mPGC), epigenetic reprogramming and pluripotency, but its role in the evolutionarily divergent regulatory network of human PGCs (hPGCs) remains unclear. Besides, a previous knockdown study indicated that PRDM14 might be dispensable for human germ cell fate. Here, we decided to use inducible degrons for a more rapid and comprehensive PRDM14 depletion. We show that PRDM14 loss results in significantly reduced specification efficiency and an aberrant transcriptome of human PGC-like cells (hPGCLCs) obtained in vitro from human embryonic stem cells (hESCs). Chromatin immunoprecipitation and transcriptomic analyses suggest that PRDM14 cooperates with TFAP2C and BLIMP1 to upregulate germ cell and pluripotency genes, while repressing WNT signalling and somatic markers. Notably, PRDM14 targets are not conserved between mouse and human, emphasising the divergent molecular mechanisms of PGC specification. The effectiveness of degrons for acute protein depletion is widely applicable in various developmental contexts.
2019-12-08 | GSE138674 | GEO
Project description:PRDM14 is a crucial regulator of mouse primordial germ cells (mPGC), epigenetic reprogramming and pluripotency, but its role in the evolutionarily divergent regulatory network of human PGCs (hPGCs) remains unclear. Besides, a previous knockdown study indicated that PRDM14 might be dispensable for human germ cell fate. Here, we decided to use inducible degrons for a more rapid and comprehensive PRDM14 depletion. We show that PRDM14 loss results in significantly reduced specification efficiency and an aberrant transcriptome of human PGC-like cells (hPGCLCs) obtained in vitro from human embryonic stem cells (hESCs). Chromatin immunoprecipitation and transcriptomic analyses suggest that PRDM14 cooperates with TFAP2C and BLIMP1 to upregulate germ cell and pluripotency genes, while repressing WNT signalling and somatic markers. Notably, PRDM14 targets are not conserved between mouse and human, emphasising the divergent molecular mechanisms of PGC specification. The effectiveness of degrons for acute protein depletion is widely applicable in various developmental contexts.
2019-12-08 | GSE138673 | GEO
Project description:The molecular mechanisms of human primordial germ cell (PGC) specification are poorly understood due to the inaccessibility of cell materials and the lack of an alternative in vitro model that enables tracking of the earliest stages of germ cell development. Here, we introduce a defined and efficient differentiation system for the induction of pre-migratory PGC-like cells from human embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs). By step-wise differentiation, we generated an OCT4+/ T+/BLIMP1+ cell population that transitioned into STELLA expressing PGC-like cells that exhibited a similar key gene expression as mouse PGCs as well as global epigenetic reprogramming. Even though, these PGC-like cells expressed PRDM14 at very low levels, they underwent activation of pluripotency/PGC genes, suppression of neural induction and suppression of de novo DNA methylation, events that are regulated by Prdm14 during mouse PGC specification. This study demonstrates that human PGC commitment shares many key features with mouse PGC specification, but harbors unique and so far unknown mechanisms that, point to a novel human transcriptional regulation.
2015-04-06 | GSE53498 | GEO
Project description:The molecular mechanisms of human primordial germ cell (PGC) specification are poorly understood due to the inaccessibility of cell materials and the lack of an alternative in vitro model that enables tracking of the earliest stages of germ cell development. Here, we introduce a defined and efficient differentiation system for the induction of pre-migratory PGC-like cells from human embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs). By step-wise differentiation, we generated an OCT4+/ T+/BLIMP1+ cell population that transitioned into STELLA expressing PGC-like cells that exhibited a similar key gene expression as mouse PGCs as well as global epigenetic reprogramming. Even though, these PGC-like cells expressed PRDM14 at very low levels, they underwent activation of pluripotency/PGC genes, suppression of neural induction and suppression of de novo DNA methylation, events that are regulated by Prdm14 during mouse PGC specification. This study demonstrates that human PGC commitment shares many key features with mouse PGC specification, but harbors unique and so far unknown mechanisms that, point to a novel human transcriptional regulation. 7 samples were analyzed. ESC: Human Embryonic Stem Cells, 1 biological rep iPSC: Human induced Pluripotent Stem Cells, 1 biological rep d2: Human induced Pluripotent Stem Cells, 2 days differentiation treatment , 2 biological rep d4PGCLC: Human induced Pluripotent Stem Cells, 4 days differentiation treatment towards Primordial Germ Cells Like Cells, 1 biological rep d6PGCLC: Human induced Pluripotent Stem Cells, 6 days differentiation treatment towards Primordial Germ Cells Like Cells, 2 biological rep
2015-04-06 | E-GEOD-53498 | biostudies-arrayexpress