Figure 4.

Identification of RIT1-RGL3– and p53-MDM2–binding domains by DoMY-Seq.A, integrative Genomics Viewer (IGV) snapshots of the RIT1-interacting domain in RGL3. Note that by using two different bait vectors and reporter systems, we obtained the same interaction motif, which is depicted below and maps within the RGL3 Ras association (RA) domain. For each assay, two biological independent assays were performed. B, similar snapshots depict the p53-binding motifs of MDM2 using two fragment size prey libraries. We observed two peaks that map to the canonical N-terminal p53 binding domain and the acidic transactivating region. Given the presence of acidic 9-aa TAD sequences, this motif can indicate a previously described binding domain or a false-positive transactivating sequence. For each assay, two biological independent assays were performed. DoMY-Seq, Protein Domain mapping using Yeast 2 Hybrid-Next Generation Sequencing; MDM2, mouse double minute 2 homolog; TAD, transactivation domain.