Figure 4.

Cellular proteins bind to the predicted Core α-MoRF regions. Single or double residue substitutions were introduced into fragments containing Core MoRF1 (A) and 2 (B) by error-prone PCR. The effect of these substitutions on binding to cellular proteins was then assessed using the Y2H assay. Substitutions in the predicted α-MoRF regions (grey blocks) preferentially disrupted binding. pOAD-IF is a control plasmid, in which URA3 is in frame with the GAL4 activation domain and no human gene is present. –TLUH indicates Y2H selection medium lacking tryptophan, leucine, uracil, and histidine; –TL, growth control medium lacking tryptophan and leucine. All interactions were assessed in independent duplicate experiments; representative results are shown in this study.