title: An aspartyl cathepsin, CTH3, is essential for proprotein processing during secretory granule maturation in Tetrahymena thermophila.
abstract: In Tetrahymena thermophila, peptides secreted via dense-core granules, called mucocysts, are generated by proprotein processing. We used expression profiling to identify candidate processing enzymes, which localized as cyan fluorescent protein fusions to mucocysts. Of note, the aspartyl cathepsin Cth3p plays a key role in mucocyst-based secretion, since knockdown of this gene blocked proteolytic maturation of the entire set of mucocyst proproteins and dramatically reduced mucocyst accumulation. The activity of Cth3p was eliminated by mutation of two predicted active-site mutations, and overexpression of the wild-type gene, but not the catalytic-site mutant, partially rescued a Mendelian mutant defective in mucocyst proprotein processing. Our results provide the first direct evidence for the role of proprotein processing in this system. Of interest, both localization and the CTH3 disruption phenotype suggest that the enzyme provides non-mucocyst-related functions. Phylogenetic analysis of the T. thermophila cathepsins, combined with prior work on the role of sortilin receptors in mucocyst biogenesis, suggests that repurposing of lysosomal enzymes was an important step in the evolution of secretory granules in ciliates.
authors.affiliation: Department of Molecular Genetics and Cell Biology, University of Chicago, Chicago, IL 60637.
authors.forename: Santosh
authors.initials: S
authors.lastname: Kumar
authors.affiliation: Department of Molecular Genetics and Cell Biology, University of Chicago, Chicago, IL 60637.
authors.forename: Joseph S
authors.initials: JS
authors.lastname: Briguglio
authors.affiliation: Department of Molecular Genetics and Cell Biology, University of Chicago, Chicago, IL 60637 apturkew@midway.uchicago.edu.
authors.forename: Aaron P
authors.initials: AP
authors.lastname: Turkewitz
mesh_terms: D002268:Carboxypeptidases / Q000378:metabolism*; D002403:Cathepsins / Q000235:genetics / Q000378:metabolism*; D005089:Exocytosis; D009154:Mutation; D010802:Phylogeny; D022142:Secretory Vesicles / Q000378:metabolism*; D016808:Tetrahymena thermophila / Q000166:cytology* / Q000235:genetics
chemical_list: D002268:Carboxypeptidases; D002403:Cathepsins