Turnover of F1F0-ATP synthase subunit 9 and other proteolipids in normal and Batten disease fibroblasts.

Fibroblasts derived from patients with late infantile neuronal ceroid lipofucsinosis (NCL) and from a mouse model of NCL are similar to cells in intact animals in that they accumulate subunit 9 of mitochondrial F1F0-ATP synthase (F-ATPase) (Tanner, A., Dice, J.F., Cell Biol. Int. 19 (1995) 71-75). We now report no differences in the synthetic rates of F-ATPase subunit 9 in such affected cells when compared to control cells. However, the degradation rates of F-ATPase subunit 9 are reduced in both the affected human and mouse cells. This reduced degradation applies only to subunit 9 and the homologous vacuolar ATPase subunit among five distinct, reproducible proteolipid bands analyzed. Approximately 15% of newly synthesized F-ATPase subunit 9 is rapidly degraded in control cells, but this rapidly degraded component is absent in both the human and mouse NCL fibroblasts. At confluence, when the accumulated F-ATPase subunit 9 transiently disappears from human NCL fibroblasts, there is an increased degradation of all proteolipids. The pathway of degradation that is enhanced at confluence is likely to correspond to lysosomal macroautophagy. We confirmed that lysosomes were able to degrade F-ATPase subunit 9 after endocytosis of radiolabeled mitochondria. Human NCL fibroblasts were less active than control cells in this lysosomal degradation of endocytosed F-ATPase subunit 9. However, this difference was not specific for F-ATPase subunit 9 since it also applied to total endocytosed mitochondrial protein. We conclude that degradation of F-ATPase subunit 9 can occur by multiple pathways and that a mitochondrial pathway of proteolysis is defective in the late infantile human and mouse forms of NCL.