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Genome-wide analysis of the role of FOG-1 in GATA-1 chromatin occupancy

ABSTRACT: We report ChIP-Seq data for GATA-1 and the FOG-binding mutant of GATA-1 (GATA-1^V205G) in G1ME cells, a Gata1-null cell line with both erythroid and megakaryocytic differentiation potential. We introduced HA-tagged GATA-1 or V205G into G1ME cells via retroviral transduction. The cells were crosslinked at 48h post-transduction, and an HA antibody was used for chromatin immunoprecipitation (ChIP). ChIP and input samples were sequenced on Illumina GAII high-throughput sequencer. The data reveal GATA-1-specific and V205G-specific bidning sites, indicating that FOG-1 both faacilitates and prohibits GATA-1 chromatin occupancy in a context-dependent manner.

ORGANISM(S): Mus musculus

PROVIDER: GSE35644 | GEO | 2012/07/30

SECONDARY ACCESSION(S): PRJNA155881

REPOSITORIES: GEO

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Genome-wide analysis of the role of FOG-1 in GATA-1 chromatin occupancy

Project description:We report ChIP-Seq data for GATA-1 and the FOG-binding mutant of GATA-1 (GATA-1^V205G) in G1ME cells, a Gata1-null cell line with both erythroid and megakaryocytic differentiation potential. We introduced HA-tagged GATA-1 or V205G into G1ME cells via retroviral transduction. The cells were crosslinked at 48h post-transduction, and an HA antibody was used for chromatin immunoprecipitation (ChIP). ChIP and input samples were sequenced on Illumina GAII high-throughput sequencer. The data reveal GATA-1-specific and V205G-specific bidning sites, indicating that FOG-1 both faacilitates and prohibits GATA-1 chromatin occupancy in a context-dependent manner. Examinaton of chromatin occupancy of GATA-1 anda FOG-binding mutant of GATA-1 in G1ME cells cultured in TPO.

2012-07-30 | E-GEOD-35644 | biostudies-arrayexpress

The role of FOG-1 in GATA-1 transcriptional regulation of megakaryocyte differentiation

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The role of FOG-1 in GATA-1 transcriptional regulation of megakaryocyte differentiation

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Genome-wide analysis of chromatin occupancy of GATA1 and its short isoform GATA1s (ChIP-Seq)

Project description:We report ChIP-Seq data for GATA1 and the leukemia-associated short isoform GATA1s in G1ME cells, a Gata1-null cell line with both erythroid and megakaryocytic differentiation potential. We introduced HA-tagged GATA1 or GATA1s into G1ME cells via retroviral transduction. The cells were crosslinked at 48h post-transduction, and an HA antibody was used for chromatin immunoprecipitation (ChIP). ChIP and input samples were sequenced on Illumina GAII or GAIIx high-throughput sequencers. The data reveal a deficiency of chromatin occupancy by GATA1s, especially at genes involved in erythrocyte differentiation and function.

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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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Genome-wide analysis of chromatin occupancy of GATA1 and its short isoform GATA1s (ChIP-Seq)

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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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