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 major types of clathrin adaptors 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 superresolution 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 (PI4P) levels at the TGN slow or uncouple AP-1 coat assembly from Gga coat assembly. Conversely, enhanced PI4P synthesis shortens the time between adaptor waves. Gga2p binds directly to the TGN PI4-kinase Pik1p and contributes to Pik1p recruitment. These results identify a PI4P-based mechanism for regulating progressive assembly of adaptor-specific clathrin coats at the TGN.