title: Identification of Cell Cycle Dependent Interaction Partners of the Septins by Quantitative Mass Spectrometry.
abstract: The septins are a conserved family of GTP-binding proteins that, in the baker's yeast, assemble into a highly ordered array of filaments at the mother bud neck. These filaments undergo significant structural rearrangements during the cell cycle. We aimed at identifying key components that are involved in or regulate the transitions of the septins. By combining cell synchronization and quantitative affinity-purification mass-spectrometry, we performed a screen for specific interaction partners of the septins at three distinct stages of the cell cycle. A total of 83 interaction partners of the septins were assigned. Surprisingly, we detected DNA-interacting/nuclear proteins and proteins involved in ribosome biogenesis and protein synthesis predominantly present in alpha-factor arrested that do not display an assembled septin structure. Furthermore, two distinct sets of regulatory proteins that are specific for cells at S-phase with a stable septin collar or at mitosis with split septin rings were identified. Complementary methods like SPLIFF and immunoprecipitation allowed us to more exactly define the spatial and temporal characteristics of selected hits of the AP-MS screen.
authors.affiliation: Ulm University, Department of Molecular Genetics and Cell Biology, Ulm, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Renz
authors.affiliation: University of Freiburg, Department of Biochemistry and Functional Proteomics, Institute of Biology II, Faculty of Biology and BIOSS Centre for Biological Signalling Studies, Freiburg, Germany.
authors.forename: Silke
authors.initials: S
authors.lastname: Oeljeklaus
authors.affiliation: Ulm University, Department of Molecular Genetics and Cell Biology, Ulm, Germany.
authors.forename: Sören
authors.initials: S
authors.lastname: Grinhagens
authors.affiliation: University of Freiburg, Department of Biochemistry and Functional Proteomics, Institute of Biology II, Faculty of Biology and BIOSS Centre for Biological Signalling Studies, Freiburg, Germany.
authors.forename: Bettina
authors.initials: B
authors.lastname: Warscheid
authors.affiliation: Ulm University, Department of Molecular Genetics and Cell Biology, Ulm, Germany.
authors.forename: Nils
authors.initials: N
authors.lastname: Johnsson
authors.affiliation: Ulm University, Department of Molecular Genetics and Cell Biology, Ulm, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Gronemeyer
mesh_terms: D001665:Binding Sites; D002453:Cell Cycle / Q000235:genetics*; D018797:Cell Cycle Proteins / Q000235:genetics* / Q000378:metabolism; D003599:Cytoskeleton / Q000235:genetics / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D013058:Mass Spectrometry / Q000379:methods; D011485:Protein Binding; D025941:Protein Interaction Mapping; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D055503:Protein Multimerization; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism; D058112:Septins / Q000235:genetics* / Q000378:metabolism
chemical_list: D018797:Cell Cycle Proteins; D020033:Protein Isoforms; D029701:Saccharomyces cerevisiae Proteins; D058112:Septins