O-GlcNAc homeostasis contributes to cell fate decisions during hematopoiesis

• Published in: The Journal of biological chemistry Vol. 294; no. 4; pp. 1363 - 1379
• Main Authors: Zhang, Zhen, Parker, Matthew P., Graw, Stefan, Novikova, Lesya V., Fedosyuk, Halyna, Fontes, Joseph D., Koestler, Devin C., Peterson, Kenneth R., Slawson, Chad
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 25-01-2019; American Society for Biochemistry and Molecular Biology
• Subjects: Acetylglucosamine - metabolism; Cell Differentiation; Cells, Cultured; differentiation; Erythroid Cells - cytology; Erythroid Cells - metabolism; erythropoiesis; FOG-1; G1E-ER4; GATA transcription factor; GATA1 Transcription Factor - metabolism; Gene Regulation; Hematopoiesis; Homeostasis; Humans; Myeloid Cells - cytology; Myeloid Cells - physiology; myelopoiesis; N-Acetylglucosaminyltransferases - metabolism; O-GlcNAcylation; O-linked N-acetylglucosamine (O-GlcNAc) transferase (OGT); post-translational modification; transcription; hematopoiesis; differentiation; transcription; gene regulation; GATA transcription factor; myelopoiesis; O-linked N-acetylglucosamine (O-GlcNAc) transferase (OGT); O-GlcNAcylation; post-translational modification; erythropoiesis; G1E-ER4; FOG-1
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.RA118.005993

The addition of a single β-d-GlcNAc sugar (O-GlcNAc) by O-GlcNAc-transferase (OGT) and O-GlcNAc removal by O-GlcNAcase (OGA) maintain homeostatic O-GlcNAc levels on cellular proteins. Changes in protein O-GlcNAcylation regulate cellular differentiation and cell fate decisions, but how these changes affect erythropoiesis, an essential process in blood cell formation, remains unclear. Here, we investigated the role of O-GlcNAcylation in erythropoiesis by using G1E-ER4 cells, which carry the erythroid-specific transcription factor GATA-binding protein 1 (GATA-1) fused to the estrogen receptor (GATA-1–ER) and therefore undergo erythropoiesis after β-estradiol (E2) addition. We observed that during G1E-ER4 differentiation, overall O-GlcNAc levels decrease, and physical interactions of GATA-1 with both OGT and OGA increase. RNA-Seq–based transcriptome analysis of G1E-ER4 cells differentiated in the presence of the OGA inhibitor Thiamet-G (TMG) revealed changes in expression of 433 GATA-1 target genes. ChIP results indicated that the TMG treatment decreases the occupancy of GATA-1, OGT, and OGA at the GATA-binding site of the lysosomal protein transmembrane 5 (Laptm5) gene promoter. TMG also reduced the expression of genes involved in differentiation of NB4 and HL60 human myeloid leukemia cells, suggesting that O-GlcNAcylation is involved in the regulation of hematopoietic differentiation. Sustained treatment of G1E-ER4 cells with TMG before differentiation reduced hemoglobin-positive cells and increased stem/progenitor cell surface markers. Our results show that alterations in O-GlcNAcylation disrupt transcriptional programs controlling erythropoietic lineage commitment, suggesting a role for O-GlcNAcylation in regulating hematopoietic cell fate.