Figure 3.
Targeted ubiquitination of LexA-Cdk2 by aptamer-hect fusions. (A) Design of a “modifier,” inspired by the structure of hect domain containing ubiquitin ligases. (B) Western blot analysis of LexA-Cdk2 (Upper) and TrxA-hect or aptamer-hect fusions (Lower) using anti-LexA and anti-TrxA antibodies, respectively. (C) Western blot analysis of LexA-Cdk2 when aptamer-hect fusions are expressed by growth overnight in a medium that does or does not contain CuSO4 and that does or does not express Myc-tagged ubiquitin. (D) Western blot analysis of LexA-Cdk2 and LexA-7Lys-Cdk2 when TrxA-, 8- and 10M-hect fusions are expressed, using the anti-LexA antibody.