Figure 3.
Assembly of the OXPHOS complexes in mutants devoid of ribosome recycling (Rrf1) or translation initiation factors (Ifm1 or Mif3). (A) Total proteins from [35S]-labeling experiments were analyzed by Western blotting with antibodies against Cox1, Cytb, Atp9, Cox2; Porin was used as a loading control (upper panel). Quantification was done using the Image J software (lower panel). (B) Cytochrome absorption spectra were recorded on whole cells. The position of the absorption maxima of cytochromes a + a3 (complex IV), cytochrome b and c1 (complex III) and c are indicated. (C and D) Mitochondrial proteins were solubilized with either dodecylmaltoside (DM, panel C) or digitonin (DG, panel D), separated on BN-PAGE 4–16% (left) or 3–12% (right) and immunoblotted with antibodies against Cyt1, Cox2 and Atp2. Positions of respiratory complexes III and IV, of supercomplexes III2 + IV2, III2 + IV as well as those of multimers (Vn), monomers (V) and F1-part of complex V are indicated. The protein molecular mass markers are also indicated (kDa).