title: Involvement of Golgi-associated retrograde protein complex in the recycling of the putative Dnf aminophospholipid flippases in yeast.
abstract: It is widely accepted that phosphatidylethanolamine (PE) is enriched in the cytosolic leaflet of the eukaryotic plasma membranes. To identify genes involved in the establishment and regulation of the asymmetric distribution of PE on the plasma membrane, we screened the deletion strain collection of the yeast Saccharomyces cerevisiae for hypersensitive mutants to the lantibiotic peptide Ro09-0198 (Ro) that specifically binds to PE on the cell surface and inhibits cellular growth. Deletion mutants of VPS51, VPS52, VPS53, and VPS54 encoding the components of Golgi-associated retrograde protein (GARP) complex, YPT6 encoding a Rab family small GTPase that functions with GARP complex, RIC1 and RGP1 encoding its guanine nucleotide exchange factor (GEF), and TLG2 encoding t-SNARE exhibited hypersensitivity to Ro. The mutants deleted for VPS51, VPS52, VPS53, and VPS54 were impaired in the uptake of fluorescently labeled PE. In addition, aberrant intracellular localization of the EGFP-tagged Dnf2p, the putative inward-directed phospholipid translocase (flippase) of the plasma membrane, was observed in the mutant defective in the GARP complex, Ypt6p, its GEF proteins, or Tlg2p. Our results suggest that the GARP complex is involved in the recycling of Dnf flippases.
authors.affiliation: Department of Biotechnology, The University of Tokyo, Tokyo, Japan.
authors.forename: Keiko
authors.initials: K
authors.lastname: Takagi
authors.forename: Kunihiko
authors.initials: K
authors.lastname: Iwamoto
authors.forename: Shingo
authors.initials: S
authors.lastname: Kobayashi
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Horiuchi
authors.forename: Ryouichi
authors.initials: R
authors.lastname: Fukuda
authors.forename: Akinori
authors.initials: A
authors.lastname: Ohta
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000235:genetics / Q000378:metabolism*; D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism*; D002462:Cell Membrane / Q000201:enzymology; D004252:DNA Mutational Analysis; D025141:Drug Resistance, Fungal; D017353:Gene Deletion; D006056:Golgi Apparatus / Q000235:genetics / Q000378:metabolism*; D010455:Peptides / Q000494:pharmacology; D010456:Peptides, Cyclic; D010714:Phosphatidylethanolamines / Q000378:metabolism*; D049294:Phospholipid Transfer Proteins / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D018528:ATP-Binding Cassette Transporters; D010455:Peptides; D010456:Peptides, Cyclic; D010714:Phosphatidylethanolamines; D049294:Phospholipid Transfer Proteins; D029701:Saccharomyces cerevisiae Proteins; C055996:lanthiopeptin; C483858:phosphatidylethanolamine; D000251:Adenosine Triphosphatases; C480602:Dnf2 protein, S cerevisiae