FIGURE 2.

Functional complementation of yeast mutants lacking the respective ATP-independent protease by plant homolog. (A) Yeast deletion mutants were transformed with plasmid carrying a full-length coding sequence of a plant protease or a coding sequence of yeast protease as a positive control. Ten-fold serial dilutions of yeast cells were spotted onto the plates with rich medium containing 2% glycerol (YPG) and incubated at indicated temperature for 3 days. Wild-type and yeast deletion mutants carrying empty plasmid were spotted as a control (vector). (B) atp23Δ yeast mutants were transformed with plasmid carrying either plant AtATP23 or yeast ATP23 gene (positive control). Mitochondria from the yeast cells were probed with anti-Atp6 antibodies. p, precursor form of Atp6; m, mature form of Atp6; “–” untransformed atp23Δ cells. (C) Yeast deletion mutants were transformed with plasmid carrying a full-length coding sequence of a plant protease. Plate assay as in (A).