EDAG positively regulates erythroid differentiation and modifies GATA1 acetylation through recruiting p300

Wei Wei Zheng, Xiao Ming Dong, Rong Hua Yin, Fei Fei Xu, Hong Mei Ning, Mei Jiang Zhang, Cheng Wang Xu, Yang Yang, Ya Li Ding, Zhi Dong Wang, Wen Bo Zhao, Liu Jun Tang, Hui Chen, Xiao Hui Wang, Yi Qun Zhan, Miao Yu, Chang Hui Ge, Chang Yan Li, Xiao Ming Yang

Research output: Contribution to journalArticlepeer-review

25 Scopus citations


Erythroid differentiation-associated gene (EDAG) has been considered to be a transcriptional regulator that controls hematopoietic cell differentiation, proliferation, and apoptosis. The role of EDAG in erythroid differentiation of primary erythroid progenitor cells and in vivo remains unknown. In this study, we found that EDAG is highly expressed in CMPs and MEPs and upregulated during the erythroid differentiation of CD34+ cells following erythropoietin (EPO) treatment. Overexpression of EDAG induced erythroid differentiation of CD34+ cells in vitro and in vivo using immunodeficient mice. Conversely, EDAG knockdown reduced erythroid differentiation in EPO-treated CD34+ cells. Detailed mechanistic analysis suggested that EDAG forms complex with GATA1 and p300 and increases GATA1 acetylation and transcriptional activity by facilitating the interaction between GATA1 and p300. EDAG deletion mutants lacking the binding domain with GATA1 or p300 failed to enhance erythroid differentiation, suggesting that EDAG regulates erythroid differentiation partly through forming EDAG/GATA1/p300 complex. In the presence of the specific inhibitor of p300 acetyltransferase activity, C646, EDAG was unable to accelerate erythroid differentiation, indicating an involvement of p300 acetyltransferase activity in EDAG-induced erythroid differentiation. ChIP-PCR experiments confirmed that GATA1 and EDAG co-occupy GATA1-targeted genes in primary erythroid cells and in vivo. ChIP-seq was further performed to examine the global occupancy of EDAG during erythroid differentiation and a total of 7,133 enrichment peaks corresponding to 3,847 genes were identified. Merging EDAG ChIP-Seq and GATA1 ChIP-Seq datasets revealed that 782 genes overlapped. Microarray analysis suggested that EDAG knockdown selectively inhibits GATA1-activated target genes. These data provide novel insights into EDAG in regulation of erythroid differentiation.

Original languageEnglish (US)
Pages (from-to)2278-2289
Number of pages12
Issue number8
StatePublished - Aug 2014

All Science Journal Classification (ASJC) codes

  • General Medicine

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