title: Characterization of Septin Ultrastructure in Budding Yeast Using Electron Tomography.
abstract: Septins are essential for the completion of cytokinesis. In budding yeast, Saccharomyces cerevisiae, septins are located at the bud neck during mitosis and are closely connected to the inner plasma membrane. In vitro, yeast septins have been shown to self-assemble into a variety of filamentous structures, including rods, paired filaments, bundles, and rings (Bertin et al. Proc Natl Acad Sci U S A, 105(24):8274-8279, 2008; Garcia et al. J Cell Biol, 195(6):993-1004, 2011; Bertin et al. J Mol Biol, 404(4):711-731, 2010). Using electron tomography of freeze-substituted sections and cryo-electron tomography of frozen sections, we determined the three-dimensional organization of the septin cytoskeleton in dividing budding yeast with molecular resolution (Bertin et al. Mol Biol Cell, 23(3):423-432, 2012; Bertin and Nogales. Commun Integr Biol 5(5):503-505, 2012). Here, we describe the detailed procedures used for our characterization of the septin cellular ultrastructure.
authors.affiliation: Biochemistry, Biophysics and Structural Biology Division, Department of Molecular and Cell Biology, University of California, Berkeley, CA, 94720, USA.|Institut Curie, CNRS UMR 168, 75231, Paris, France.
authors.forename: Aurélie
authors.initials: A
authors.lastname: Bertin
authors.affiliation: Biochemistry, Biophysics and Structural Biology Division, Department of Molecular and Cell Biology, University of California, Berkeley, CA, 94720, USA. enogales@lbl.gov.|Howard Hughes Medical Institute, University of California, 708C Stanley Hall, Berkeley, CA, 94720-3220, USA. enogales@lbl.gov.|Life Sciences Division, Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Berkeley, CA, 94720, USA. enogales@lbl.gov.
authors.forename: Eva
authors.initials: E
authors.lastname: Nogales
mesh_terms: D020285:Cryoelectron Microscopy / Q000379:methods; D048749:Cytokinesis / Q000235:genetics; D055032:Electron Microscope Tomography / Q000379:methods*; D007091:Image Processing, Computer-Assisted; D004718:Saccharomycetales / Q000648:ultrastructure*; D058112:Septins / Q000737:chemistry*
chemical_list: D058112:Septins
keywords: Budding yeast; Cryo-sectio ning; Cryo-tomography; Cytokinesis; Image process ing; Septin