Protein Arginine Methyltransferase 5 Functions in Opposite Ways in the Cytoplasm and Nucleus of Prostate Cancer Cells
Figure 4
The PRMT5-p44 interaction is essential for the PRMT5-promoted p44 cytoplasmic localization.
(A) Mutations in p44 abolished PRMT5-driven p44 cytoplasmic translocation. Cells were transfected with pcDNA-GFP-p44(WT) or pcDNA-GFP-p44(MT) alone or together with pcDNA-PRMT5. The nucleus was stained with Far-red, and the subcellular localization of GFP-p44 was observed under a confocal microscope. (B) Mutations in p44 abolished the interaction of p44 with PRMT5. Cells were transfected with pcDNA–f:p44 (WT) (lane 1) or pcDNA–f:p44 (MT) (lanes 2–5), and whole-cell lysates were prepared for immunoprecipitation with anti-FLAG antibody (M2 agarose). Western blot with anti-PRMT5 was performed to detect the precipitated PRMT5 (bottom panel). Top panel shows expression of wild-type (WT) or mutated (MT) p44 in the lysates used for the immunoprecipitation.