Figure 2.

Selection for reporter activation by Gal4–sfGFP–Fis1p reveals TA mutations inhibiting mitochondrial localization. (A) Missense mutations within the Fis1p TA provide selectable marker activation. Strain MaV203 expressing Gal4–sfGFP–Fis1p from plasmid b100 or variants expressed from plasmids b128 (V145E), b129 (L139P), b130 (L129P, V138A), or b101 (∆TA) were treated as in Figure 1C. pKS1 (vector) is also provided as a negative control. (B) Missense mutations within the Fis1p TA allow cytosolic and nuclear accumulation of a linked mCherry protein. mCherry fused to variants of the Fis1p TA were expressed in WT strain CDD961 from plasmids b109 (WT), b134 (V145E), b135 (L139P), b136 (L129P, V138A), or b252 (∆TA) and visualized by fluorescence microscopy. Mitochondria were labeled with a mitochondria-targeted GFP expressed from plasmid pHS1. Bar, 5 µm.