Yeast Cytokinesis pp 113-123

Part of the Methods in Molecular Biology book series (MIMB, volume 1369)

Characterization of Septin Ultrastructure in Budding Yeast Using Electron Tomography


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.

Key words

Septin Budding yeast Cytokinesis Cryo-tomography Image processing Cryo-sectioning 


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Biochemistry, Biophysics and Structural Biology Division, Department of Molecular and Cell BiologyUniversity of CaliforniaBerkeleyUSA
  2. 2.Institut Curie, CNRS UMR 168ParisFrance
  3. 3.Howard Hughes Medical InstituteUniversity of CaliforniaBerkeleyUSA
  4. 4.Life Sciences DivisionLawrence Berkeley National LaboratoryBerkeleyUSA

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