Cancer-associated mutations inhibit DHHC20 function.
A, endogenous DHHC20, F-actin (arrows, top panels), and MCAM (bottom panels) localization. B, asymmetric DHHC20 localization is independent of Wnt5a (n > 300 cells/experimental condition; Student's t test; **, p < 0.005). Error bars, S.D. C, levels of palmitoylated MCAM decrease in WM239A cells expressing DHHC20 shRNA compared with control shRNA. D, positions of DHHC15 mutations (black) and analogous amino acids in DHHC20 (gray). E, HEK293T cells expressing DHHC20 shRNA were transfected with MCAM-GFP and wild type (WT) DHHC20 or mutants C156S, K151Q, and P157S that are shRNA-resistant. Cells were labeled with palmitic acid azide. Empty vector (EV) and unlabeled wild type cells (UL/WT). F, immunoprecipitation of wild type and mutant DHHC20-CFP-FLAG probed with anti-MCAM. G, representative cells expressing YFP-tagged DHHC20 wild type and point mutants in WM239A melanoma cells.