title: Over-expression of EDAG in the myeloid cell line 32D: induction of GATA-1 expression and erythroid/megakaryocytic phenotype.
abstract: Erythroid differentiation-associated gene (EDAG), a hematopoietic tissue-specific transcription regulator, plays a key role in maintaining the homeostasis of hematopoietic lineage commitment. However, the mechanism and genes regulated by EDAG remain unknown. In this study, we showed that overexpression of EDAG in a myeloid cell line 32D led to an erythroid phenotype with increased number of benzidine-positive cells and up-regulation of erythroid specific surface marker TER119. The megakaryocytic specific marker CD61 was also induced significantly. Using a genome-wide microarray analysis and a twofold change cutoff, we identified 332 genes with reduced expression and 288 genes with increased expression. Among up-regulation genes, transcription factor GATA-1 and its target genes including EKLF, NF-E2, Gfi-1b, hemogen, SCL, hemoglobin alpha, beta and megakaryocytic gene GPIX were increased. Silencing of EDAG by RNA interference in K562 cells resulted in down-regulation of these genes. Taken together, EDAG functions as a positive regulator of erythroid/megakaryocytic differentiation in 32D cells associated with the induction of GATA-1 and its target genes.
authors.affiliation: Beijing Institute of Radiation Medicine, Beijing 100850, China.
authors.forename: Ya-Li
authors.initials: YL
authors.lastname: Ding
authors.forename: Cheng-Wang
authors.initials: CW
authors.lastname: Xu
authors.forename: Zhi-Dong
authors.initials: ZD
authors.lastname: Wang
authors.forename: Yi-Qun
authors.initials: YQ
authors.lastname: Zhan
authors.forename: Wei
authors.initials: W
authors.lastname: Li
authors.forename: Wang-Xiang
authors.initials: WX
authors.lastname: Xu
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: D002454:Cell Differentiation; D004912:Erythrocytes / Q000166:cytology / Q000378:metabolism*; D005434:Flow Cytometry; D050982:GATA1 Transcription Factor / Q000235:genetics*; D020868:Gene Silencing; D006801:Humans; D007377:Interleukin-3 / Q000378:metabolism; D020014:K562 Cells; D008533:Megakaryocytes / Q000166:cytology / Q000378:metabolism*; D009687:Nuclear Proteins / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis; D010641:Phenotype; D034622:RNA Interference; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation
chemical_list: D050982:GATA1 Transcription Factor; C494688:GATA1 protein, human; C435745:HEMGN protein, human; D007377:Interleukin-3; D009687:Nuclear Proteins