title: Partial genetic suppression of a loss-of-function mutant of the neuronal ceroid lipofuscinosis-associated protease TPP1 in Dictyostelium discoideum.
abstract: Neuronal ceroid lipofuscinosis (NCL) is the most common childhood-onset neurodegenerative disease. NCL is inevitably fatal, and there is currently no treatment available. Children with NCL show a progressive decline in movement, vision and mental abilities, and an accumulation of autofluorescent deposits in neurons and other cell types. Late-infantile NCL is caused by mutations in the lysosomal protease tripeptidyl peptidase 1 (TPP1). TPP1 cleaves tripeptides from the N-terminus of proteins in vitro, but little is known about the physiological function of TPP1. TPP1 shows wide conservation in vertebrates but it is not found in Drosophila, Caenorhabditis elegans or Saccharomyces cerevisiae. Here, we characterize ddTpp1, a TPP1 ortholog present in the social amoeba Dictyostelium discoideum. Lysates from cells lacking ddTpp1 show a reduced but not abolished ability to cleave a TPP1 substrate, suggesting that other Dictyostelium enzymes can perform this cleavage. ddTpp1 and human TPP1 localize to the lysosome in Dictyostelium, indicating conserved function and trafficking. Cells that lack ddTpp1 show precocious multicellular development and a reduced ability to form spores during development. When cultured in autophagy-stimulating conditions, cells lacking ddTpp1 rapidly decrease in size and are less viable than wild-type cells, suggesting that one function of ddTpp1 could be to limit autophagy. Cells that lack ddTpp1 exhibit strongly impaired development in the presence of the lysosome-perturbing drug chloroquine, and this phenotype can be suppressed through a secondary mutation in the gene that we name suppressor of tpp1(-) A (stpA), which encodes a protein with some similarity to mammalian oxysterol-binding proteins (OSBPs). Taken together, these results suggest that targeting specific proteins could be a viable way to suppress the effects of loss of TPP1 function.
authors.affiliation: Department of Biology, Texas A&M University, College Station, TX 77843-3474, USA.
authors.forename: Jonathan E
authors.initials: JE
authors.lastname: Phillips
authors.affiliation: Department of Biology, Texas A&M University, College Station, TX 77843-3474, USA. rgomer@tamu.edu.
authors.forename: Richard H
authors.initials: RH
authors.lastname: Gomer
mesh_terms: D000626:Aminopeptidases / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D001343:Autophagy; D004023:Dictyostelium / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / Q000235:genetics* / Q000378:metabolism; D005453:Fluorescence; D016153:Genes, Suppressor; D006801:Humans; D008247:Lysosomes / Q000378:metabolism; D009154:Mutation / Q000235:genetics*; D009472:Neuronal Ceroid-Lipofuscinoses / Q000201:enzymology*; D021381:Protein Transport; D059748:Proteolysis; D057057:Serine Proteases / Q000235:genetics* / Q000378:metabolism; D013170:Spores / Q000254:growth & development; D013489:Suppression, Genetic; D013997:Time Factors; D000091346:Tripeptidyl-Peptidase 1
chemical_list: D000091346:Tripeptidyl-Peptidase 1; D057057:Serine Proteases; D000626:Aminopeptidases; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases; C000717267:TPP1 protein, human
keywords: Batten disease; Dictyostelium; Neuronal ceroid lipofuscinosis; TPP1; Tripeptidyl peptidase 1