Figure 5.
The intermediate form of Ccp1 (iCcp1) is not enriched at the IBM of mitochondria. (A) Expression of s-Mgm1* partially suppresses the respiratory growth defect of Δpcp1 cells. Tenfold serial dilutions of logarithmically growing cultures were spotted onto agar plates containing glucose or glycerol as sole carbon source and incubated for 7 d at 30°C. (B) Δpcp1 cells exhibit mitochondria with aberrant internal structures. Frequently, mitochondria without cristae (left) or with onion-shaped membrane inclusions (right) were observed. (C) The mitochondria of Δpcp1 cells expressing s-Mgm1* frequently contain cristae. Exemplary EM image of a mitochondrion of a Δpcp1 cells expressing s-Mgm1* immunogold labeled for Ccp1-GFP; the arrowheads point to gold particles. Right, quantification of the distribution of iCcp1-GFP revealed an enrichment (68% of all gold particles) of iCcp1-GFP within the mitochondrial interior. (D) Live-cell fluorescence microscopy of both Ccp1-GFP and the CM-marker Qcr2-mRFP in Δpcp1 Δmdm10 cells. Single confocal sections are displayed. Inset, normalized intensity profiles along the indicated area. Scale bars, (B and C) 200 nm; (D) 3 μm.