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. 2000 Sep 12;97(20):10757–10762. doi: 10.1073/pnas.190062797

Figure 4.

Figure 4

The LxxLL motifs are critical for LZIP activation function. (A) Wild-type and mutant versions of Gal4-LZIPN92 were cotransfected into BHK21 cells together with the p5xGal-E1B-luc reporter (500 ng), and luciferase activity was measured after 40 h. (B) Yeast two-hybrid interaction assay to monitor association between the β-propeller domain of HCF-1 and wild-type or mutant LZIPN92 fragments. A yeast GAL1-LacZ reporter strain was transformed with expression plasmids encoding the wild-type or P134S mutant HCF-1 β-propeller domain (residues 1–380) fused to the LexA DNA-binding domain together with plasmids expressing wild-type, LxxLL 1KO, LxxLL 2KO, LxxLL1/2KO, and LxxLLHBMKO versions of LZIPN92 fused to the B42 activation domain. Strains were grown on 5-bromo-4-chloro-3-indolyl β-D-galactoside indicator plates. For each combination, five independent transformants were patched.