Methods for evaluating the Caenorhabditis elegans dauer state: standard dauer-formation assay using synthetic daumones and proteomic analysis of O-GlcNAc modifications

Methods Cell Biol. 2011;106:445-60. doi: 10.1016/B978-0-12-544172-8.00016-5.

Abstract

The dauer state is a non-feeding, alternative L3 state characterized by a number of distinctive metabolic and morphological changes. There are many naturally occurring dauer-inducing pheromones, termed daumones, that have been suggested by some to exhibit differences in dauer-inducing activity. Here, we have established a standard dauer-formation assay that uses synthetic daumones 1, 2, and 3, the three major daumones. To analyze the proteome of Caenorhabditis elegans in the dauer state, we focused on O-GlcNAc modification, a cytosolic modification of proteins that is known to interact either competitively or synergistically with protein phosphorylation. Protein O-GlcNAc modification is an important biological process in cells that can ensure the timely response to extracellular stimuli, such as daumone, and maintain cellular homeostasis. Establishing a standard method for assaying dauer formation using different synthetic daumones, and using differences in O-GlcNAcylated proteins during the dauer state to analyze the dauer proteome will lead to a better understanding of dauer biology of C. elegans in the context of animal longevity and adaptation under harsh environments.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Animals
  • Caenorhabditis elegans / metabolism
  • Caenorhabditis elegans / physiology*
  • Caenorhabditis elegans Proteins / chemistry
  • Caenorhabditis elegans Proteins / metabolism*
  • Culture Techniques
  • Electrophoresis, Gel, Two-Dimensional
  • Fatty Acids / metabolism*
  • Fatty Acids / pharmacology
  • Fatty Acids / physiology
  • Glycosylation
  • Pheromones / metabolism*
  • Pheromones / pharmacology
  • Pheromones / physiology
  • Protein Processing, Post-Translational
  • Proteolysis
  • Proteomics
  • Tandem Mass Spectrometry

Substances

  • Caenorhabditis elegans Proteins
  • Fatty Acids
  • Pheromones
  • daumone, C elegans