Fig. 1. Nuclear export of YAN is mediated by CRM1 and blocked by insertion of a NLS into YAN. (A-C,E-L) S2 cultured cells transfected with various YAN constructs and stained with anti-YAN. (A'-C',E'-L') DAPI staining of the same cells. (D) Schematic of YAN showing predicted domains and sites of SV40 Large T-antigen NLS insertions. For each experiment (A-C,E-L), the percentage of transfected cells exhibiting nuclear localization (A,C,E-K) or exclusively cytoplasmic localization (B,L) is indicated. n, number of cells scored in each experiment. (A-C') YAN^WT; (E-F') YAN^{Int NLS}; (G-H') YAN^{2x NLS}; (I-J') YAN^{N' NLS}; (K-L') YAN^{Mut NLS}. (A,E,G,I,K) YAN localization in the absence of RAS^V12 (B,F,H,J,L) YAN localization in the presence of RAS^V12. (C) YAN localization in the presence of RAS^V12 and RNAi of crm1. (C) YAN localization is restricted to the nucleus in the presence of RAS^V12 and RNAi of crm1. (F,H) Internal NLS insertions completely inhibit nuclear export of YAN in the presence of RAS^V12, while the N-terminal insertion only partially prevents export (J). (L) Insertion of a nonfunctional NLS into YAN has no effect on export.

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