Fig. 1.

INA complex deficiency affects biogenesis of mitochondrial-encoded F₁F₀-ATP synthase subunits. a Schematic presentation of the INA complex. IMS, intermembrane space; N, N-terminus, C, C-terminus. b Mitochondrial-encoded proteins were radiolabeled in vivo and analyzed by urea SDS-PAGE and digital autoradiography. Atp6 and Cox1 signals obtained after a 20 min pulse were normalized to Cox3 signal and protein amounts in mutant mitochondria were quantified relative to wild type (wild type = 100%), (n = 3, ±SEM). c Mitochondrial-encoded proteins were radiolabeled in vivo for 10 min. After labeling, samples were chased with an excess of methionine for 30 or 90 min, and analyzed as in b. Atp6 and Cox3 signals after 90 min chase were quantified as the percentage of their starting signals (0 min time point), (n = 3, ±SEM). d Mitochondrial-encoded proteins were radiolabeled in organello for 20 min and analyzed by SDS-PAGE and autoradiography. To quantify Atp9 levels, samples were TCA-precipitated prior to SDS-PAGE and autoradiography. After normalization to Cox1, Atp8, Atp9, Cox3/Atp6, and cytochrome b (Cytb) signals in ina22Δ and ina17Δ mitochondria were quantified as the percentage of wild type (n = 3, ±SEM)