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. 2004 Aug 26;23(18):3643–3652. doi: 10.1038/sj.emboj.7600370

Figure 2.

Figure 2

Permutations of selected peptides mediate distinct CRM1-mediated subcellular localisation. (A) Amino-acid sequences of P0, P1, S0 and S1 15-mer peptides compared to the HIV-1 Rev NES and the NES consensus sequence. NES sequences are in bold and consensus hydrophobic amino acids (φ) are underlined. One-letter amino-acid abbreviations are used. (B) Shuttling reporter proteins containing GFP, the export-incompetent Rev(1.4) variant and peptide sequences from (A) were transiently expressed in MCF7 cells and subcellular localisation was detected by green fluorescence 24 h post-transfection. The effect of 50 nM LMB was monitored 3 h after addition. (C) Identification of the critical amino-acid residues in S1 NES. Positions 3 and 8 in P0 and S1 NESs were interchanged (left), and subcellular localisation of the GFP reporter plasmids was analysed as in (B) (right). (D) Mutagenesis of the natural RanBP1 NES. A peptide corresponding to the natural RanBP1 NES was mutated to conform the high-affinity NES consensus (left). Subcellular localisation was analysed as above (right).