title: Protein kinase Gin4 negatively regulates flippase function and controls plasma membrane asymmetry.
abstract: Plasma membrane function requires distinct leaflet lipid compositions. Two of the P-type ATPases (flippases) in yeast, Dnf1 and Dnf2, translocate aminoglycerophospholipids from the outer to the inner leaflet, stimulated via phosphorylation by cortically localized protein kinase Fpk1. By monitoring Fpk1 activity in vivo, we found that Fpk1 was hyperactive in cells lacking Gin4, a protein kinase previously implicated in septin collar assembly. Gin4 colocalized with Fpk1 at the cortical site of future bud emergence and phosphorylated Fpk1 at multiple sites, which we mapped. As judged by biochemical and phenotypic criteria, a mutant (Fpk1(11A)), in which 11 sites were mutated to Ala, was hyperactive, causing increased inward transport of phosphatidylethanolamine. Thus, Gin4 is a negative regulator of Fpk1 and therefore an indirect negative regulator of flippase function. Moreover, we found that decreasing flippase function rescued the growth deficiency of four different cytokinesis mutants, which suggests that the primary function of Gin4 is highly localized control of membrane lipid asymmetry and is necessary for optimal cytokinesis.
authors: Division of Biochemistry, Biophysics and Structural Biology, Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720.
authors: Françoise M
authors: FM
authors: Roelants
authors: Division of Biochemistry, Biophysics and Structural Biology, Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720.
authors: Brooke M
authors: BM
authors: Su
authors: Division of Biochemistry, Biophysics and Structural Biology, Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720.
authors: Joachim
authors: J
authors: von Wulffen
authors: Division of Biochemistry, Biophysics and Structural Biology, Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720.
authors: Subramaniam
authors: S
authors: Ramachandran
authors: Division of Biochemistry, Biophysics and Structural Biology, Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720.
authors: Elodie
authors: E
authors: Sartorel
authors: Division of Biochemistry, Biophysics and Structural Biology, Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720.
authors: Amy E
authors: AE
authors: Trott
authors: Division of Biochemistry, Biophysics and Structural Biology, Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720 jthorner@berkeley.edu.
authors: Jeremy
authors: J
authors: Thorner
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000378:metabolism*; D000251:Adenosine Triphosphatases / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism* / Q000648:ultrastructure; D018844:Cyclin-Dependent Kinases / Q000502:physiology*; D048749:Cytokinesis; D008563:Membrane Lipids / Q000378:metabolism; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D021381:Protein Transport; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000201:enzymology*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism* / Q000502:physiology*
chemical_list: D018528:ATP-Binding Cassette Transporters; D008563:Membrane Lipids; D029701:Saccharomyces cerevisiae Proteins; D011494:Protein Kinases; C533049:Fpk1 protein, S cerevisiae; C107283:GIN4 protein, S cerevisiae; D018844:Cyclin-Dependent Kinases; D000251:Adenosine Triphosphatases; C480602:Dnf2 protein, S cerevisiae; C480601:Dnf1 protein, S cerevisiae
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