E2s interacting with intact RING finger domain of Cbl.

(A) Representative yeast colonies transformed with wild-type Cbl or RF Mutant Cbl C381A as the bait and with the indicated E2s as prey. Empty DNA-binding domain (DB) and empty activation domain (AD) vectors were used as negative controls for bait and prey, respectively. (B) Representative yeast colonies transformed with activated Cbl Y371E or activated RF mutant Cbl Y371E/C381A as the bait with the indicated E2s as prey. Ube2e family members and Ube2w interact with activated Cbl Y371E but not with activated RF mutant Cbl Y371E/C381A. For both A and B, the left panel shows growth of the yeast on -His, -Leu, -Trp, +3AT interaction selective media and the right panel shows grow on -Leu, -Trp media to confirm that the yeast received both bait and prey plasmids.