Phosphoinositide-mediated clathrin adaptor progression at the trans-Golgi network
Copyright policy )Phosphoinositide-mediated clathrin adaptor progression at the trans-Golgi network
Clathrin-coated vesicles mediate endocytosis and transport between the trans-Golgi network (TGN) and endosomes in eukaryotic cells. Clathrin adaptors play central roles in coat assembly, interacting with clathrin, cargo and membranes. Two main types of clathrin adaptor act in TGN-endosome traffic: GGA proteins and the AP-1 complex. Here we characterize the relationship between GGA proteins, AP-1 and other TGN clathrin adaptors using live-cell and super-resolution microscopy in yeast. We present evidence that GGA proteins and AP-1 are recruited sequentially in two waves of coat assembly at the TGN. Mutations that decrease phosphatidylinositol 4-phosphate (PtdIns(4)P) levels at the TGN slow or uncouple AP-1 coat assembly from GGA coat assembly. Conversely, enhanced PtdIns(4)P synthesis shortens the time between adaptor waves. Gga2p binds directly to the TGN PtdIns(4)-kinase Pik1p and contributes to Pik1p recruitment. These results identify a PtdIns(4)P-based mechanism for regulating progressive assembly of adaptor-specific clathrin coats at the TGN.
- University of California, Los Angeles United States
- University of California, San Francisco United States
- University of California, Los Angeles
- University of California United States
- University of California United States
Saccharomyces cerevisiae Proteins, 1.1 Normal biological development and functioning, Adaptor Protein Complex 1, Immunoblotting, Endosomes, Saccharomyces cerevisiae, Medical and Health Sciences, Time-Lapse Imaging, Fluorescence, Article, Phosphatidylinositol Phosphates, Underpinning research, 1-Phosphatidylinositol 4-Kinase, Microscopy, Microscopy, Confocal, Adaptor Proteins, Clathrin-Coated Vesicles, Biological Sciences, Clathrin, Vesicular Transport, Adaptor Proteins, Vesicular Transport, Luminescent Proteins, Protein Transport, Microscopy, Fluorescence, Confocal, Mutation, Generic health relevance, Developmental Biology, Protein Binding, trans-Golgi Network
Saccharomyces cerevisiae Proteins, 1.1 Normal biological development and functioning, Adaptor Protein Complex 1, Immunoblotting, Endosomes, Saccharomyces cerevisiae, Medical and Health Sciences, Time-Lapse Imaging, Fluorescence, Article, Phosphatidylinositol Phosphates, Underpinning research, 1-Phosphatidylinositol 4-Kinase, Microscopy, Microscopy, Confocal, Adaptor Proteins, Clathrin-Coated Vesicles, Biological Sciences, Clathrin, Vesicular Transport, Adaptor Proteins, Vesicular Transport, Luminescent Proteins, Protein Transport, Microscopy, Fluorescence, Confocal, Mutation, Generic health relevance, Developmental Biology, Protein Binding, trans-Golgi Network
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