Histone demethylase JMJD3 regulates cell cycle-related gene expression pattern on eargly stage of human erythropoiesis [CUT&Tag]
The Jumonji domain-containing protein D3 (JMJD3) is a histone demethylases that specifically demethylate di-and tri-methyl-lysine 27 on histone H3 (H3K27me2/3) to regulate correlation gene expression. JMJD3 promotes the differentiation and apoptosis of leukemia cells, and inhibits cell proliferation by regulating the methylation level of H3K27. JMJD3 promotes Zebrafish myeloid development and inhibits erythroid development. However, the function and mechanism of JMJD3 in human erythropoiesis still remains largely unknown. Here, we demonstrated that JMJD3 was expressed throughout the erythroid development process, with higher expression in the early stages. During the early erythropoiesis, JMJD3 deficiency caused cell cycle arrest in the G0/G1 phase, which impaired cell growth and erythroid progenitors differentiation, but did not result in cell apoptosis. Cut&tag sequencing and RNA sequencing analysis discovered that JMJD3 regulated the expression of cell cycle and erythrocyte differentiation associated factor lymphoid enhancer-binding factor 1(LEF1) dependently of H3K27me3 modifications. Our findings provide novel insights into understanding of the roles of JMJD3 and H3K27me3 in the regulation of erythropoiesis, and further indicates the significance of epigenetic regulation for the erythropoiesis. Overall design: The JMJD3/UTX inhibitor GSK-J4 (Catalog #HY-5648B; MedChemExpress) was dissolved in DMSO and administered to cell cultures at a final concentration of 1 and 1.5 µM. After treated with 1µM DZNep for 5 days, we performed a H3K27me3 CUT&Tag sequencing of DMSO and GSK-J4 treated cells. ?