Fig. 4: Characterization of Emf1 as a mitochondria component.

a C6_03200W (renamed EMF1) is an essential C. albicans gene. The tetO-EMF1/emf1∆ strain was grown and assessed for essentiality as described in Fig. 3a. b A co-expression network for EMF1 identifies multiple mitochondrial proteins in the top 50 co-expressed genes. Nodes represent genes, and edges represent the strength of the co-expression. All genes had a co-expression score of at least 0.997. Green indicates mitochondrial annotation, light blue indicates translation annotation, gray indicates no GO term annotation available. c Depletion of EMF1 perturbs mitochondrial morphology. Strains were grown overnight in the absence or presence of 0.05 µg/mL DOX, subcultured to an OD₆₀₀ of 0.1 with the same respective DOX conditions, and grown for 3 h at 30 °C. Cultures were further incubated with 50 nM Mitotracker Red for 40 m, washed, and resuspended in PBS. MitoTracker Red staining was imaged with the DsRed channel with equal exposure among samples. Experiment was performed in biological duplicate with similar results. d Emf1 localizes to the mitochondria. Cells were subcultured to an OD₆₀₀ of 0.1 and allowed to grow for 4 h before visualization. As indicated, cultures were incubated with 50 nM Mitotracker Red for 40 m, washed, and resuspended in PBS. Emf1 (green) and mitochondria (MitoTracker, red) were visualized by fluorescence microscopy. Experiment was performed in biological duplicate with similar results. e Emf1 co-localizes with Gcf1 at DAPI-stained mitochondrial nucleoids. Cells were subcultured to an OD₆₀₀ of 0.1 and grown for 4 h before being washed and resuspended in PBS, then incubated with 1 µg/mL DAPI for 1 h. Emf1 (green), Gcf1 (red), and DNA (DAPI, blue) were visualized by fluorescence microscopy. Experiment was performed in biological duplicate with similar results. f Transcriptional repression of EMF1 causes a significant reduction in mtDNA copy number. Relative NAD2 copy number in wild-type, tetO-EMF1/emf1∆, and tetO-GCF1/gcf1∆ cells in the absence and presence of 0.05 µg/mL DOX as determined by qPCR, using ACT1 and GPD1 for normalization. Values shown are relative NAD2 copy number compared to the wild-type strain in the absence of DOX. Error bars represent SEM above and below the mean of technical triplicates (two-way ANOVA, Bonferroni correction for multiple comparisons, **P < 0.002; ***P < 0.0004; ****P < 0.0001 compared to wild-type untreated). Experiment was performed in biological duplicate with similar results. Source data are provided as a Source Data file.