FIGURE 9.

Autoinhibition of Mycd WT/CRM1 interaction by N-terminal region of Mycd WT (Mycd N128). A, inhibitory effect of Mycd N128 on the interaction between CRM1 and Mycd ΔN128. The CRM1-binding affinities of Mycd ΔN128 and Mycd ΔN128/ΔCB were examined in the absence or presence of Mycd N128. Mixtures of HA-tagged CRM1, GTP-bound GST-FLAG-RanQ69L, and each indicated FLAG-tagged Mycd were subjected to IP analyses as described in the legend for Fig. 1. The respective IP/IB signal intensities were quantified as described under “Experimental Procedures.” The percentages at the top of the gels indicate relative binding affinities for CRM1 normalized by the respective affinities of Mycd ΔN128 and Mycd ΔN128/ΔCB in the absence of Mycd N128 (−), which were set at 100%. Results are means ± S.E. of three independent experiments. B, effect of the CB deletion on the Mycd WT/CRM1 interaction. Mixtures of HA-tagged CRM1, GTP-bound GST-FLAG-RanQ69L, and each indicated FLAG-tagged Mycd were subjected to IP analyses. The respective IP/IB signal intensities were quantified. The percentages at the top of the gels indicate relative binding affinities for CRM1 normalized by the affinity of Mycd WT, which was set at 100%. Results are means ± S.E. of three independent experiments.