title: Golgi-dependent transport of vacuolar sorting receptors is regulated by COPII, AP1, and AP4 protein complexes in tobacco.
abstract: The cycling of vacuolar sorting receptors (VSRs) between early and late secretory pathway compartments is regulated by signals in the cytosolic tail, but the exact pathway is controversial. Here, we show that receptor targeting in tobacco (Nicotiana tabacum) initially involves a canonical coat protein complex II-dependent endoplasmic reticulum-to-Golgi bulk flow route and that VSR-ligand interactions in the cis-Golgi play an important role in vacuolar sorting. We also show that a conserved Glu is required but not sufficient for rate-limiting YXX-mediated receptor trafficking. Protein-protein interaction studies show that the VSR tail interacts with the μ-subunits of plant or mammalian clathrin adaptor complex AP1 and plant AP4 but not that of plant and mammalian AP2. Mutants causing a detour of full-length receptors via the cell surface invariantly cause the secretion of VSR ligands. Therefore, we propose that cycling via the plasma membrane is unlikely to play a role in biosynthetic vacuolar sorting under normal physiological conditions and that the conserved Ile-Met motif is mainly used to recover mistargeted receptors. This occurs via a fundamentally different pathway from the prevacuolar compartment that does not mediate recycling. The role of clathrin and clathrin-independent pathways in vacuolar targeting is discussed.
authors.affiliation: Centre for Plant Sciences, School of Biology, Faculty of Biological Sciences, University of Leeds, Leeds LS2 9JT, United Kingdom.
authors.forename: David C
authors.initials: DC
authors.lastname: Gershlick
authors.forename: Carine de Marcos
authors.initials: Cde M
authors.lastname: Lousa
authors.forename: Ombretta
authors.initials: O
authors.lastname: Foresti
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Lee
authors.forename: Estela A
authors.initials: EA
authors.lastname: Pereira
authors.forename: Luis L P
authors.initials: LL
authors.lastname: daSilva
authors.forename: Francesca
authors.initials: F
authors.lastname: Bottanelli
authors.forename: Jurgen
authors.initials: J
authors.lastname: Denecke
mesh_terms: D000595:Amino Acid Sequence; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism*; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D021381:Protein Transport; D014026:Nicotiana / Q000378:metabolism*; D014617:Vacuoles / Q000378:metabolism*
chemical_list: D010940:Plant Proteins