title: An endosomal syntaxin and the AP-3 complex are required for formation and maturation of candidate lysosome-related secretory organelles (mucocysts) inTetrahymena thermophila.
abstract: The ciliateTetrahymena thermophilasynthesizes large secretory vesicles called mucocysts. Mucocyst biosynthesis shares features with dense core granules (DCGs) in animal cells, including proteolytic processing of cargo proteins during maturation. However, other molecular features have suggested relatedness to lysosome-related organelles (LROs). LROs, which include diverse organelles in animals, are formed via convergence of secretory and endocytic trafficking. Here we analyzedTetrahymenasyntaxin 7-like 1 (Stx7l1p), a Qa-SNARE whose homologues in other lineages are linked with vacuoles/LROs. Stx7l1p is targeted to both immature and mature mucocysts and is essential in mucocyst formation. InSTX7L1-knockout cells, the two major classes of mucocyst cargo proteins localize independently, accumulating in largely nonoverlapping vesicles. Thus initial formation of immature mucocysts involves heterotypic fusion, in which a subset of mucocyst proteins is delivered via an endolysosomal compartment. Further, we show that subsequent maturation requires AP-3, a complex widely implicated in LRO formation. Knockout of the µ-subunit gene does not impede delivery of any known mucocyst cargo but nonetheless arrests mucocyst maturation. Our data argue that secretory organelles in ciliates may represent a new class of LROs and reveal key roles of an endosomal syntaxin and AP-3 in the assembly of this complex compartment.
authors.affiliation: Department of Molecular Genetics and Cell Biology, University of Chicago, Chicago, IL 60637.
authors.forename: Harsimran
authors.initials: H
authors.lastname: Kaur
authors.affiliation: Department of Molecular Genetics and Cell Biology, University of Chicago, Chicago, IL 60637.
authors.forename: Daniela
authors.initials: D
authors.lastname: Sparvoli
authors.affiliation: Advanced ICT Research Institute, National Institute of Information and Communications Technology, Kobe 651-2492, Japan.
authors.forename: Hiroko
authors.initials: H
authors.lastname: Osakada
authors.affiliation: Advanced ICT Research Institute, National Institute of Information and Communications Technology, Kobe 651-2492, Japan.
authors.forename: Masaaki
authors.initials: M
authors.lastname: Iwamoto
authors.affiliation: Advanced ICT Research Institute, National Institute of Information and Communications Technology, Kobe 651-2492, Japan.|Graduate School of Frontier Biosciences, Osaka University, Suita 565-0871, Japan.
authors.forename: Tokuko
authors.initials: T
authors.lastname: Haraguchi
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: D000818:Animals; D001692:Biological Transport / Q000502:physiology; D011992:Endosomes / Q000378:metabolism; D005089:Exocytosis / Q000235:genetics; D008247:Lysosomes / Q000378:metabolism; D015388:Organelles / Q000378:metabolism; D021381:Protein Transport / Q000502:physiology; D015800:Protozoan Proteins / Q000378:metabolism; D050765:Qa-SNARE Proteins / Q000378:metabolism*; D022142:Secretory Vesicles / Q000378:metabolism*; D016808:Tetrahymena thermophila / Q000235:genetics / Q000378:metabolism*
chemical_list: D015800:Protozoan Proteins; D050765:Qa-SNARE Proteins