title: EDAG positively regulates erythroid differentiation and modifies GATA1 acetylation through recruiting p300.
abstract: 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.
authors.affiliation: State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Beijing, People's Republic of China.
authors.forename: Wei-Wei
authors.initials: WW
authors.lastname: Zheng
authors.forename: Xiao-Ming
authors.initials: XM
authors.lastname: Dong
authors.forename: Rong-Hua
authors.initials: RH
authors.lastname: Yin
authors.forename: Fei-Fei
authors.initials: FF
authors.lastname: Xu
authors.forename: Hong-Mei
authors.initials: HM
authors.lastname: Ning
authors.forename: Mei-Jiang
authors.initials: MJ
authors.lastname: Zhang
authors.forename: Cheng-Wang
authors.initials: CW
authors.lastname: Xu
authors.forename: Yang
authors.initials: Y
authors.lastname: Yang
authors.forename: Ya-Li
authors.initials: YL
authors.lastname: Ding
authors.forename: Zhi-Dong
authors.initials: ZD
authors.lastname: Wang
authors.forename: Wen-Bo
authors.initials: WB
authors.lastname: Zhao
authors.forename: Liu-Jun
authors.initials: LJ
authors.lastname: Tang
authors.forename: Hui
authors.initials: H
authors.lastname: Chen
authors.forename: Xiao-Hui
authors.initials: XH
authors.lastname: Wang
authors.forename: Yi-Qun
authors.initials: YQ
authors.lastname: Zhan
authors.forename: Miao
authors.initials: M
authors.lastname: Yu
authors.forename: Chang-Hui
authors.initials: CH
authors.lastname: Ge
authors.forename: Chang-Yan
authors.initials: CY
authors.lastname: Li
authors.forename: Xiao-Ming
authors.initials: XM
authors.lastname: Yang
mesh_terms: D000107:Acetylation; D000818:Animals; D015153:Blotting, Western; D002454:Cell Differentiation / Q000502:physiology*; D002469:Cell Separation; D050881:E1A-Associated p300 Protein / Q000378:metabolism*; D041905:Erythroid Cells / Q000166:cytology / Q000378:metabolism; D005260:Female; D005434:Flow Cytometry; D050982:GATA1 Transcription Factor / Q000378:metabolism*; D006410:Hematopoiesis / Q000502:physiology*; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D064593:Heterografts; D006801:Humans; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D009687:Nuclear Proteins / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D059467:Transcriptome
chemical_list: D050982:GATA1 Transcription Factor; C494688:GATA1 protein, human; C435745:HEMGN protein, human; D009687:Nuclear Proteins; D050881:E1A-Associated p300 Protein; C494360:EP300 protein, human
keywords: Acetylation; Erythroid differentiation; Erythroid differentiation-associated gene; GATA1; p300