title: Isolation and characterization of novel mutations in CDC50, the non-catalytic subunit of the Drs2p phospholipid flippase.
abstract: Flippases (type 4 P-type ATPases) are believed to translocate phospholipids from the exoplasmic to the cytoplasmic leaflet in bilayer membranes. Since flippases are structurally similar to ion-transporting P-type ATPases such as the Ca(2+) ATPase, one important question is how flippases have evolved to transport phospholipids instead of ions. We previously showed that a conserved membrane protein, Cdc50p, is required for the endoplasmic reticulum exit of the Drs2p flippase in yeast. However, Cdc50p is still associated with Drs2p after its transport to the endosomal/trans-Golgi network (TGN) membranes, and its function in the complex with Drs2p is unknown. In this study, we isolated novel temperature-sensitive (ts) cdc50 mutants whose products were still localized to endosomal/TGN compartments at the non-permissive temperature. Mutant Cdc50 proteins colocalized with Drs2p in endosomal/TGN compartments, and they co-immunoprecipitated with Drs2p. These cdc50-ts mutants exhibited defects in vesicle transport from early endosomes to the TGN as the cdc50 deletion mutant did. These results suggest that mutant Cdc50 proteins could be complexed with Drs2p, but the resulting Cdc50p-Drs2p complex is functionally defective at the non-permissive temperature. Cdc50p may play an important role for phospholipid translocation by Drs2p.
authors.affiliation: Division of Molecular Interaction, Institute for Genetic Medicine, Hokkaido University, N15 W7, Kita-ku, Sapporo, 060-0815, Japan.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Takahashi
authors.forename: Konomi
authors.initials: K
authors.lastname: Fujimura-Kamada
authors.forename: Satoshi
authors.initials: S
authors.lastname: Kondo
authors.forename: Kazuma
authors.initials: K
authors.lastname: Tanaka
mesh_terms: D000252:Calcium-Transporting ATPases / Q000235:genetics*; D009154:Mutation; D010743:Phospholipids / Q000378:metabolism; D021122:Protein Subunits / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics*
chemical_list: C427246:CDC50 protein, S cerevisiae; C104014:DRS2 protein, S cerevisiae; D010743:Phospholipids; D021122:Protein Subunits; D029701:Saccharomyces cerevisiae Proteins; D000252:Calcium-Transporting ATPases