Exploring the Functional Complementation between Grp94 and Hsp90
Fig 3
Select Grp94 domains support yeast viability.
(A) Plasmid shuffle experiments demonstrate grpN-hspMC and hspNM1grpM2C are able to support yeast viability when expressed as the sole Hsp90. Cells were plated on FOA plates as 5-fold serial dilutions and were grown at the indicated temperatures for 3โ9 days. Images are from day 4. Green highlights indicate positive complementation. (B) Western blot of yeast whole cell lysates. 20 ฮผg of total protein extracted from the lysis of plasmid-transformed yeast cells grown in SDโUra -Trp was separated by SDS-PAGE. The expression level of plasmid-expressed hsp90 chimeras was evaluated by Western blotting against the N-terminal His6 epitope tag. Lysate from Chimera 1 (yeast Hsp90) was included on each blot as a transfer and antibody control. Coomassie blue stained PVDF membranes used for blotting are displayed below their respective luminographs to allow comparison of total protein levels.
