GSE161608
ChIP seq for GATA-1 and HDAC1 in G1E cells with wild type GATA-1 or mutant
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. Overall design: ChIP seq for GATA-1 and HDAC1 in G1E cells with wild type GATA-1 or mutant