FIGURE 1.

Y14 interacts with PRMT5 and the cytoplasmic methylosome. A, HeLa cell cytoplasmic extract was subjected to a pulldown assay using GST or GST-Y14/His-Magoh heterodimer (YM) as bait. Pulled down proteins were detected by silver staining of SDS-polyacrylamide gels (upper panel) and immunoblotting with anti-PRMT5 (lower panel). Lane M, molecular mass markers. B, HeLa cell cytoplasmic (Cyto.) or nuclear (Nucl.) extract was subjected to GST pulldown assays as in A. Immunoblotting was performed using antibodies against PRMT5, pICln, and MEP50. C, the pulldown assay was performed as in A. The bait proteins were as indicated above the gel. For both B and C, the bait proteins were detected by Coomassie (Cooma.) Brilliant Blue staining (lower panels). A schematic diagram shows the domain structure of Y14 and its truncated versions. RRM, RNA recognition motif; a.a., amino acids.