Figure 5.—
Inactivation of ubl-5 perturbs formation of protein complexes in the mitochondria. (A) Detection of an ∼75-kDa biotinylated protein(s) in fractionated worm extracts by strepavidin–HRP ligand blotting. Total worm extract (T), postmitochondrial supernatant (SN), and mitochondrial pellet (MT) are shown. GFPcyt expressed from the myo-3 promoter serves as a marker for a cytoplasmic protein. (B) Distribution of an ∼75-kDa biotinylated mitochondrial protein(s) in fractions of glycerol gradient prepared from animals exposed to mock RNAi, hsp-60(RNAi), and ubl-5(RNAi). (Top) Unstressed animals. (Bottom) Mitochondrially stressed myo-3∷gfpmt animals. The migration of complexes of the indicated size in this gradient is shown. The final fraction (14) also contains the pellet.