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Embryonic erythropoiesis and hemoglobin switching require transcriptional repressor ETO2 to modulate chromatin accessibility and looping (ChIP-Seq)

ABSTRACT: To determine direct targets and the regulatory role of ETO2 in gene expression, we performed ChIPmentation with antibodies to unmodified ETO2 and ETO2 interacted factor LDB1. A de novo MEME search performed on ETO2-occupied sites, revealed enrichment for GATA and TAL binding motifs, which are the components in LDB1 complex. Nearly 86% of ETO2-binding sites were intergenic or intronic, suggesting ETO2 functions primarily in regions of the genome likely to encompass enhancers. Notabley, ETO2 occupancy was almost completely coincident with LDB1 (97% of 7,761 sites), consistent with the presence of ETO2 exclusively within the LDB1 complex and their co-function in gene regulation.

ORGANISM(S): Homo sapiens

PROVIDER: GSE142227 | GEO | 2020/08/25

REPOSITORIES: GEO

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Embryonic erythropoiesis and hemoglobin switching require transcriptional repressor ETO2 to modulate chromatin accessibility and looping (RNA-Seq)

Project description:ETO2 functions as a transcription repressor and is required for the embryonic erythropoiesis and the hemoglobin switch. To gain insight into ETO2 regulatory function during human erythropoiesis, we performed RNA-seq for WT and ETO2 KO K562 cells and found that up-regulated genes upon ETO2 loss in human cells included many markers of mature erythroid cells EPB42, ALAS2, GYPA and SLC25a37. Notably, the α-globin genes (HBA1, HBA2 and HBZ) and embryonic and fetal β-globin genes (HBE1, HBG1, and HBG2) were significantly increased after deletion of ETO2. By contrast, deletion of ETO2 down-regulated the transcription factor genes (ETS1, KLF8 and SOX6) which play a negative role in globin gene expression and hemoglobin synthesis. To further explore different domain function of ETO2, we have analyzed our RNA-seq data from domain deletion cell lines compared to the cell line expressing wild type ETO2. Generally, 702 genes were found to co-regulated by three domain function of ETO2. Interestingly, the regulation of fetal globin genes, erythroid regulator (SOX5) as well as epigenetic factor (HDAC7) is required by three domain function of ETO2. During mouse embryonic erythropoiesis, our FACS-sorted and normal RNA-seq data in E14.5 fetal liver cells indicated that eto2 promoted a critical developmental transition and played an important role in globin switch from embryonic to adult β-globin transcription since its function is essential for key regulators (PU.1, BCL11A and ZBTB7A) and globin genes (Hbb-y and Hba-x) regulation.

2020-08-25 | GSE142226 | GEO

Embryonic erythropoiesis and hemoglobin switching require transcriptional repressor ETO2 to modulate chromatin accessibility and looping [RNA-Seq 2]

Project description:During the human cord blood CD34+ cell differentiation, expression of the genes which contribute to erycyte maturation are increased, inculding ALAS2, SLC25A37, GYPA and KLF1. ETO2 functions as a transcription repressor and is required for the erythrocyte maturation and the hemoglobin switch. During mouse embryonic erythropoiesis, RNA-seq data in E8.5 yolk sac /E12.5, E14.5 fetal liver cells indicated that eto2 promoted a critical developmental transition and played an important role in globin switch from embryonic to adult β-globin transcription since its function is essential for erythorid maturation regulators (Alas2,Slc25a37,Epb42,Gypc,Klf1) and globin genes (Hbb-y and Hba-x) regulation.

2020-08-25 | GSE156306 | GEO

ChIP-Seq of CBFA2T3 (ETO2) in CD34+ HSPC and AML cells

Project description:ETO2 (CBFA2T3) functions as a transcriptional corepressor, which can directly bind to E-protein family of transcriptional factors. Translocation of ETO2 has been implicated in multiple leukemogenic pathways. ETO2 also plays an important role in self-renewal, proliferation, and expansion of hematopoietic stem/progenitor cells (HSPCs). Here we report the genomic binding sites of ETO2 in human cord-blood derived CD34+ HSPCs, Kasumi-1 t(8;21) AML cells, and U937 AML cells. All ChIP-Seq assays were performed with an ETO2 specific antibody that does not recognize other ETO/MTG family proteins (ETO or MTGR1).

2019-05-29 | GSE126953 | GEO

Embryonic erythropoiesis and hemoglobin switching require transcriptional repressor ETO2 to modulate chromatin organization.

Project description: Not available

| S-EPMC7544236 | biostudies-literature

Distinct Ldb1/NLI complexes orchestrate γ-globin repression and re-activation through ETO2 in human adult erythroid cells.

Project description:The Ldb1/GATA-1/TAL1/LMO2 complex mediates long range interaction between the β-globin locus control region (LCR) and gene in adult mouse erythroid cells but whether this complex mediates chromatin interactions at other developmental stages or in human cells is unknown. We investigated human NLI (Ldb1 homologue) complex occupancy and chromatin conformation of the β-globin locus in human erythroid cells. In addition to the LCR, we find robust NLI complex occupancy at a site downstream of the Aγ-globin gene, within sequences of BGL3, an intergenic RNA transcript. In cells primarily transcribing β-globin, BGL3 is not transcribed and BGL3 sequences are occupied by NLI core complex members together with co-repressor ETO2 and γ-globin repressor BCL11A. The LCR and β-globin gene establish proximity in these cells. In contrast, when γ-globin transcription is re-activated by cytokines in these cells, ETO2 participation in the NLI complex at BGL3 is diminished, as is BCL11A occupancy, and both BGL3 and γ-globin are transcribed. In these cells, proximity between the BGL3/γ-globin region and the LCR is established. Thus, alternative NLI complexes mediate γ-globin transcription or silencing through long range LCR interactions involving an intergenic site of non-coding RNA transcription and ETO2 is critical to this process. ChIP-chip of GATA-1, NLI, and pol II in cell lines Comparison of GATA-1, NLI, and pol II binding on chromatin. 2 replicates for each protein; ChIP and input DNA for each replicate

2012-01-05 | E-GEOD-30047 | biostudies-arrayexpress

Distinct Ldb1/NLI complexes orchestrate γ-globin repression and re-activation through ETO2 in human adult erythroid cells.

2012-01-05 | GSE30047 | GEO

Human K562 cells overexpressing ETO2

Project description:microRNA profiling of human K562 cells comparing control or ETO2-overexpressed cells Two-condition experiment, K562-pcDNA vs. K562-pcDNA-ETO2, Biological replicates: 1, 1 pcDNA, 1 pcDNA-ETO2, independently.

2014-10-01 | E-GEOD-51485 | biostudies-arrayexpress

Human K562 cells overexpressing ETO2

Project description:microRNA profiling of human K562 cells comparing control or ETO2-overexpressed cells

2014-10-01 | GSE51485 | GEO

Identification of ETO2-GLIS2 transcriptional targets

Project description:Acute megakaryoblastic leukemia (AMKL) is a subtype of leukemia primarily diagnosed in childhood and generally associated with poor prognosis. Genetic alterations found in de novo childhood AMKL include the OTT-MAL fusion, MLL and NUP98 fusions and the recently identified ETO2-GLIS2 fusion that involves two transcriptional regulators. In order to identify ETO2-GLIS2 target genes, we performed two approaches: 1-Ectopic expression of ETO2-GLIS2, ETO2, GLIS2 and OTT-MAL in HEL cells, which does not endogenously express AMKL fusion oncogenes. Transduced cells marked by expression of the GFP were sorted by flow cytometry 24 hours after transduction and RNA was extracted with the Qiagen Rneasy kit including DNase treatment. 2-Expression of a small peptide (NC128), which interferes with the dimerization and cofactor recruitment by the ETO2-GLIS2 fusion, within MO7E cells derived from an AMKL patient and expressing endogenously the ETO2-GLIS2 fusion. Transduced cells marked by expression of the GFP were sorted by flow cytometry 7 days after transduction and RNA was extracted with the Qiagen Rneasy kit including DNase treatment. After quantification of biological replicates, and quality control (Bioanalyser, Agilent), RNA were hybridized on Agilent arrays as indicated below.

2017-05-05 | E-MTAB-4332 | biostudies-arrayexpress

Human K562 cells transfected with control or ETO2 siRNA

Project description:microRNA profiling of comparing control or ETO2 siRNA-treated human K562 cells Two-condition experiment, K562-control siRNA vs. K562-ETO2 siRNA, Biological replicates: 1, 1 control siRNA, 1 ETO2 siRNA, independently.

2014-10-01 | E-GEOD-51486 | biostudies-arrayexpress

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