Figure 3.

A bipartite localization motif regulates the localization of TRAM. (a–d) Fluorescence microscopy of macrophages transfected with plasmids encoding chimeric proteins consisting of full-length TRAM (a) or various amino acids (in parentheses) of TRAM (b–d) fused in-frame to GFP. Outlined areas are enlarged in bottom left corners. Scale bars, 5 μm. Data are representative of at least three independent experiments with over 500 cells per condition, in which over 95% of the cells had similar staining patterns (unless stated otherwise elsewhere). (e) Isoelectric points of sequential 20–amino acid segments of TRAM. Data are representative of experiments done two times. (f) Primary structure of mouse TRAM. Below, bipartite localization motif (amino acids 1–20), with the myristoylation motif (red letters) followed by the polybasic motif (substituted amino acids are underlined). (g) Amino acid alignment of human proteins identified as containing a bipartite localization motif, with putative myristoylation motifs (underlined residues) followed by the polybasic motif, and with basic and aromatic residues indicated by red lettering.