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RNAseq of Wild Type and GATA-1 mutants in G1E cells

ABSTRACT: RNAseq of Wild Type and GATA-1 mutants in G1E cells

PROVIDER: PRJNA678895 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR13072081_1.fastq.gz	Fastqsanger.gz
	SRR13072081_2.fastq.gz	Fastqsanger.gz
	SRR13072082_1.fastq.gz	Fastqsanger.gz
	SRR13072082_2.fastq.gz	Fastqsanger.gz
	SRR13072083_1.fastq.gz	Fastqsanger.gz

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RNAseq of Wild Type and GATA-1 mutants in G1E cells

Project description:Purpose: To compare gene regulation changes in GATA-1 mutant compared with wild type Methods: G1E cells with wild type GATA-1 or mutant were induced by β-estrodiol Results: We mapped the sequence reads to the mouse genome (build mm10) with TopHat workflow. Some of altered expression of genes was confirmed with qRT–PCR.

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ChIP seq for GATA-1 and HDAC1 in G1E cells with wild type GATA-1 or mutant

Project description:The transcription factor GATA-1 is essential for erythroid and megakaryocytic cell differentiation and maturation. Previous reports show that GATA-1 is modulated through acetylation modification and through FOG-1 mediated indirect intereaction with HDAC1/2 containing NuRD corepressor complexes. In this study, we found that NuRD does not deacetylate GATA-1. However, HDAC1 alone can efficiently deacetylates GATA-1 and the direct interaction of HDAC1 and GATA-1 is required for the deacetylation. Two arginines within GATA-1 linker region are important for this interaction and arginine to alanine mutations (2RA mutant) largely reduces GATA-1 binding to HDAC1 in FOG-1 independent manner. GATA-1 2RA mutant were then introduced into G1E cells, a GATA-1-null erythroid progenitor cells. The 2RA mutant is acetylated but fails to induce erythroid differentiation. Gene expression analysis shows that GATA-1 2RA mutant affects GATA-1 function in both GATA-1 activated and repressed genes. The gene expression pattern partially overlap with gene expression profile of GATA-1V205M, a GATA-1 mutant with defective FOG-1 binding. ChIP-seq analysis further reveal that 2RA mutation largely reduced GATA-1 chromatin binding, most profoundly at gene promoter regions. HDAC1 recruitment on those promoters are also strongly reduced. These results revealed that GATA-1 recruits HDAC1 to GATA-1 regulated gene promoters and HDAC1 is required for GATA-1 mediated transcription regulation.

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Project description:This SuperSeries is composed of the following subset Series: GSE24333: Expression data from ETO2 knockdown G1E-ER-GATA-1 cells GSE24336: Expression data from LMO2 knockdown G1E-ER-GATA-1 cells Refer to individual Series

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Foxo3 Regulated Genes in G1E-ER-GATA-1 Cells

Project description:The ensemble of Foxo3-regulated genes in the erythroid G1E-ER-GATA-1 cell line was determined by knocking down Foxo3 using siRNA, and measuring genome wide transcription by microarray analysis G1E-ER-GATA-1 cells were treated with control or Foxo3-specific siRNA by nucleofection at t = 0 h and t = 24 h. At t = 24 h, cells were treated with M-CM-^_-estradiol to activate ER-GATA-1. RNA was harvested at t = 48 h and processed for microarray analysis.

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Project description:Identification of genes regulated by GATA-1 independent of the cofactor FOG-1. Experiment Overall Design: A conditionally activated FOG-1-binding defective mutant of GATA-1, ER-GATA-1(V205G), was expressed in GATA-1-null G1E cells. Transcipt levels were compared in cells untreated or treated with estradiol to activate the GATA-1 mutant.

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Expression data from LMO2 knockdown G1E-ER-GATA-1 cells

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