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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. `HECTPub2' and `HECT
CAPub2' 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.