title: Mammalian GGAs act together to sort mannose 6-phosphate receptors.
abstract: The GGAs (Golgi-localized, gamma ear-containing, ADP ribosylation factor-binding proteins) are multidomain proteins implicated in protein trafficking between the Golgi and endosomes. We examined whether the three mammalian GGAs act independently or together to mediate their functions. Using cryo-immunogold electron microscopy, the three GGAs were shown to colocalize within coated buds and vesicles at the trans-Golgi network (TGN) of HeLa cells. In vitro binding experiments revealed multidomain interactions between the GGAs, and chemical cross-linking experiments demonstrated that GGAs 1 and 2 form a complex on Golgi membranes. RNA interference of each GGA resulted in decreased levels of the other GGAs and their redistribution from the TGN to cytosol. This was associated with impaired incorporation of the cation-independent mannose 6-phosphate receptor into clathrin-coated vesicles at the TGN, partial redistribution of the receptor to endosomes, and missorting of cathepsin D. The morphology of the TGN was also altered. These findings indicate that the three mammalian GGAs cooperate to sort cargo and are required for maintenance of TGN structure.
authors.affiliation: Department of Internal Medicine, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: Pradipta
authors.initials: P
authors.lastname: Ghosh
authors.forename: Janice
authors.initials: J
authors.lastname: Griffith
authors.forename: Hans J
authors.initials: HJ
authors.lastname: Geuze
authors.forename: Stuart
authors.initials: S
authors.lastname: Kornfeld
mesh_terms: D020727:ADP-Ribosylation Factors / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D033942:Adaptor Proteins, Vesicular Transport; D002352:Carrier Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D002402:Cathepsin D / Q000378:metabolism; D022163:Clathrin-Coated Vesicles / Q000378:metabolism / Q000648:ultrastructure; D003600:Cytosol / Q000378:metabolism; D015536:Down-Regulation / Q000502:physiology; D011992:Endosomes / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D007150:Immunohistochemistry; D007425:Intracellular Membranes / Q000378:metabolism / Q000648:ultrastructure; D046911:Macromolecular Substances; D008854:Microscopy, Electron; D009844:Oligosaccharides / Q000378:metabolism; D021381:Protein Transport / Q000502:physiology*; D034741:RNA, Small Interfering / Q000494:pharmacology; D017527:Receptor, IGF Type 2 / Q000378:metabolism*; D021601:trans-Golgi Network / Q000378:metabolism* / Q000648:ultrastructure
chemical_list: D033942:Adaptor Proteins, Vesicular Transport; D002352:Carrier Proteins; C407306:GGA adaptor proteins; D046911:Macromolecular Substances; D009844:Oligosaccharides; D034741:RNA, Small Interfering; D017527:Receptor, IGF Type 2; D002402:Cathepsin D; D020727:ADP-Ribosylation Factors