Analysis of the Archaeal ESCRT Apparatus

Methods Mol Biol. 2019:1998:1-11. doi: 10.1007/978-1-4939-9492-2_1.

Abstract

Members of the archaeal domain of life that lack homologs of actin and tubulin divide by binary fission in a process that is dependent upon orthologs of eukaryotic ESCRT components. Many of these archaeal organisms are hyperthermophilic acidophiles with unique cell wall structures, which create technical challenges for performing traditional cell biological techniques. Here, we describe the "baby machine" method for synchronizing microorganisms at high temperatures in order to study cell cycle-related processes. We also provide details for fixing, permeabilizing, and staining archaeal cells and ESCRT assemblies for observation by light microscopy.

Keywords: Archaea; Baby machine; Cell cycle; Cell division; ESCRT; Immunofluorescence; Sulfolobus; Synchronization.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Archaeal Proteins / metabolism*
  • Cell Cycle
  • Endosomal Sorting Complexes Required for Transport / metabolism*
  • Microscopy, Fluorescence / methods
  • Sulfolobus acidocaldarius / metabolism*

Substances

  • Archaeal Proteins
  • Endosomal Sorting Complexes Required for Transport