Comparative transcriptomics reveals mechanisms underlying cln3-deficiency phenotypes in Dictyostelium

• Published in: Cellular signalling 2019-06, Vol.58, p.79-90
• Main Authors: Huber, Robert J., Mathavarajah, Sabateeshan
• Format: Article
• Language: English
• Subjects: Batten disease; CLN3; Dictyostelium discoideum; Neuronal ceroid lipofuscinosis; RNA sequencing
• ISSN: 0898-6568; 1873-3913
• DOI: 10.1016/j.cellsig.2019.02.004

Mutations in CLN3 cause a juvenile form of neuronal ceroid lipofuscinosis (NCL). This devastating neurological disorder, commonly known as Batten disease, is currently untreatable due to a lack of understanding of the physiological role of the protein. Recently, work in the social amoeba Dictyostelium discoideum has provided valuable new insight into the function of CLN3 in the cell. More specifically, research has linked the Dictyostelium homolog (gene: cln3, protein: Cln3) to protein secretion, adhesion, and aggregation during starvation, which initiates multicellular development. In this study, we used comparative transcriptomics to explore the mechanisms underlying the aberrant response of cln3− cells to starvation. During starvation, 1153 genes were differentially expressed in cln3− cells compared to WT. Among the differentially expressed genes were homologs of other human NCL genes including TPP1/CLN2, CLN5, CTSD/CLN10, PGRN/CLN11, and CTSF/CLN13. STRING and GO term analyses revealed an enrichment of genes linked to metabolic, biosynthetic, and catalytic processes. We then coupled the findings from the RNA-seq analysis to biochemical assays, specifically showing that loss of cln3 affects the expression and activity of lysosomal enzymes, increases endo-lysosomal pH, and alters nitric oxide homeostasis. Finally, we show that cln3− cells accumulate autofluorescent storage bodies during starvation and provide evidence linking the function of Cln3 to Tpp1 and CtsD activity. In total, this study enhances our knowledge of the molecular mechanisms underlying Cln3 function in Dictyostelium. •Loss of cln3 alters the expression of 1153 genes during Dictyostelium starvation.•Loss of cln3 in Dictyostelium affects the expression of homologs of NCL genes.•Loss of cln3 in Dictyostelium affects the expression and activity of lysosomal enzymes.•cln3-deficiency increases endo-lysosomal pH in Dictyostelium.•cln3-deficiency alters nitric oxide homeostasis in Dictyostelium.