Visualization of in vivo septin ultrastructures by platinum replica electron microscopy

Methods Cell Biol. 2016;136:73-97. doi: 10.1016/bs.mcb.2016.03.011. Epub 2016 Apr 16.

Abstract

Septins are cytoskeletal proteins involved in diverse biological processes including cytokinesis, cell morphogenesis, motility, and ciliogenesis. Septins form various filamentous structures in vitro and in vivo, but the higher-order architecture of septin structures in vivo remains poorly defined. The best understood system in this respect is the budding yeast Saccharomyces cerevisiae, where septins form a ring structure that undergoes multiple stages of remodeling during the cell cycle. In this chapter, we describe a method for visualizing supramolecular septin structures in yeast at high spatial resolution using platinum replica electron microscopy. This approach can be applied to further understand the regulation of assembly and remodeling of septin higher-order structures, as well as the relationship between septin architecture and function.

Keywords: Cytokinesis; Cytoskeleton; Electron microscopy; S. cerevisiae; Septin.

MeSH terms

  • Cell Cycle Proteins / chemistry
  • Cell Cycle Proteins / ultrastructure*
  • Cytokinesis / genetics
  • Microscopy, Electron / methods*
  • Multiprotein Complexes / chemistry
  • Multiprotein Complexes / ultrastructure*
  • Platinum / chemistry
  • Saccharomyces cerevisiae / ultrastructure
  • Septins / chemistry
  • Septins / ultrastructure*

Substances

  • Cell Cycle Proteins
  • Multiprotein Complexes
  • Platinum
  • Septins