LAUSR

Mutations in at least 13 different genes (called CLNs) underlie various forms of neuronal ceroid lipofuscinoses (NCLs), a group of the most common neurodegenerative lysosomal storage diseases. While inactivating mutations in the CLN1 gene, encoding palmitoyl‐protein thioesterases‐1 (PPT1), cause infantile NCL (INCL), those in the CLN3 gene, encoding a protein of unknown function, underlie juvenile NCL (JNCL). PPT1 depalmitoylates S‐palmitoylated proteins (constituents of ceroid) required for their degradation by lysosomal hydrolases and PPT1‐deficiency causes lysosomal accumulation of autofluorescent ceroid leading to INCL. Because intracellular accumulation of ceroid is a characteristic of all NCLs, a common pathogenic link for these diseases has been suggested. It has been reported that CLN3‐mutations suppress the exit of cation‐independent mannose 6‐phosphate receptor (CI‐M6PR) from the trans Golgi network (TGN). Because CI‐M6PR transports soluble proteins such as PPT1 from the TGN to the lysosome, we hypothesized that CLN3‐mutations may cause lysosomal PPT1‐insufficiency contributing to JNCL pathogenesis. Here, we report that the lysosomes in Cln3‐mutant mice, which mimic JNCL, and those in cultured cells from JNCL patients, contain significantly reduced levels of Ppt1‐protein and Ppt1‐enzyme activity and progressively accumulate autofluorescent ceroid. Furthermore, in JNCL fibroblasts the V0a1 subunit of v‐ATPase, which regulates lysosomal acidification, is mislocalized to the plasma membrane instead of its normal location on lysosomal membrane. This defect dysregulates lysosomal acidification, as we previously reported in Cln1 −/− mice, which mimic INCL. Our findings uncover a previously unrecognized role of CLN3 in lysosomal homeostasis and suggest that CLN3‐mutations causing lysosomal Ppt1‐insuffiiciency may at least in part contribute to JNCL pathogenesis.