FIGURE 2.

Complementation assays for Wt and mutant yeast VAS1 constructs. CW1, a yeast VAS1 knock-out strain, was transformed with Wt and mutant VAS1 constructs, and the ability of the transformants to rescue the growth defect of the knock-out strain was tested. A, schematic summary of VAS1 constructs and their complementation activities. Shaded boxes and dashed lines denote sequences coding for yeast ValRS and deletions, respectively. The symbols “+” and “-” indicate positive and negative complementation, respectively. Mit, mitochondrial; Cyt, cytoplasmic. B, complementation assays for cytoplasmic ValRS activity on a 5-FOA plate. C, complementation assays for mitochondrial ValRS activity on a YPG plate. D, assay of protein expression by Western blotting. Upper panel, ValRS; lower panel, PGK (as a loading control). Numbers 1–4 (circled) in B and C represent the constructs shown in A.