Fig. 7. The in vivo functional assay of a putative catalytic HECT domain of Pub2p. (A) Schematic representation of chimeric proteins of the Pub2 HECT domain fused with the N-terminal portion of Pub1p. `HECT<sup>Pub2</sup>' and `HECT CA^Pub2' indicate the HECT domain of wild-type and the Cys639 Ala mutant Pub2p, respectively. (B) Complementation of the temperature-sensitive growth of pub1 by the chimeric proteins. pREP41-based plasmids carrying each chimeric construct were transformed into the KKT81-3A strain, which was grown on thiamine-free EMM2 plates at 37°C for 3 days.