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. 2006 Jun 8;25(12):2847–2855. doi: 10.1038/sj.emboj.7601178

Figure 1.

Figure 1

Yeast two-hybrid interactions between TLS polymerases and Ub. (A) Diagram of the gene structure of UbA52 and UbB. (B) Yeast two-hybrid assay showing the interaction (by activation of the ADE2 and HIS3 reporter genes) between full-length polι and UbA52 and UbB. In these, and all subsequent two-hybrid assays, 10 μl of an overnight culture was spotted on CSM -Leu, -Trp, -His, -Ade plates and incubated at 30°C for 5 days. (C) Deletion mapping of polι reveals that the interaction with recombinant Ub is localized to the C-terminal 224 amino acids of polι. (D) In addition to polι, polη also binds to Ub. However, neither the p125 and p66 subunits of polδ bind Ub. PCNA was used as a positive control for polι, polη, and p66 of polδ. The interaction between polι and polη and Ub is specific, as neither polymerase binds to the Ub-like protein, SUMO-1. TDG was used as a positive control for SUMO-1.