Protein Arginine Methyltransferase 1 Interacts with and Activates p38α to Facilitate Erythroid Differentiation
Figure 1
Ectopic expression of PRMT1 enhanced erythroid differentiation of K562 cells.
(A) K562 cells were transiently transfected with either empty vectors or pPCDNA3HA2-PRMT1 plasmids expressing HA-PRMT1 and were treated with either araC (1 µM) or NaB (2 mM). Transient expression of PRMT1 significantly enhanced erythroid differentiation, as detected by benzidine staining. (B) R2–1 and R2–3 cells, which stably expressed HA-PRMT1, also exhibited higher erythroid differentiation. E3–6 cells were an empty vector control. (C, D) The expression of various erythroid-related genes and transcription factors was examined by real-time PCR after araC treatment. The β-actin gene was a control whose expression was not affected by araC treatment. Data were calculated as fold changes using vector control cells without treatment as a reference. (E) The enzymatically impaired G80R PRMT1 mutant, when introduced by transient transfection, failed to enhance differentiation. (F) The stable clone that expressed the G80R mutant also failed to stimulate ALAS2 gene expression. All results shown are representative of three separate experiments. Differentiation results are presented as means ± S.E. of three repeats; *, p<0.05; **, p<0.01; ***, p<0.005 compared with vector control cells.
