Fig. 1

Phe53 substitution with Leu in SDHAF3 leads to impaired function of SDHAF3 in yeast. a Respiratory growth of yeast cells expressing a human Sdh7 ortholog, SDHAF3, and its sequences variants. Pre-cultured cells in synthetic complete (SC) media were serially diluted and then spotted on SC media containing different carbon sources, as indicated. Cells were incubated at 30 °C. b Relative succinate:quinone oxidoreductase (SQR) activity in isolated mitochondria. Mitochondria were isolated from cells grown until late-log phase in SC media plus 2% raffinose/0.2% glucose. Data are shown as mean ± SD (n = 3; **p < 0.05). c Steady-state levels of SDHAF3 proteins in sdh4∆ cells. Human SDHAF3 and its mutants under yeast MET25 promoter were expressed from plasmids in cells. Por1 (porin) is a loading control. d Steady-state levels of SDHAF3 proteins in response to ectopic expression of Sdh2-His₆Myc₂. Rip1 is the Fe/S cluster subunit of bc1 complex. e Co-immunoprecipitation of SDHAF3 was performed with mitochondrial isolated from cells expressing Sdh2-His₆Myc₂ using anti-SDHAF3 antibody and protein A magnetic beads. Anti-HA antibody is a negative control. *, non-specific reactivity; Input, 1% of total lysates