O-GlcNAc Chromatin Marks from C. elegans L1 animals

Nutrient-driven O-GlcNAcylation of key components of the transcription machinery may epigenetically modulate gene expression in metazoans. Knockouts of the O-GlcNAc cycling enzymes in C. elegans are viable and fertile, allowing a global analysis of the impact of GlcNAcylation. Whole genome ChIP profiling of the O-GlcNAc cycling mutants identified over 800 genes associated with GlcNAcylated chromatin. This novel chromatin mark is preferentially found with genes involved in stress, longevity, and innate immunity, the same set of processes affected by global changes in gene expression analysis. This experiment collected data on fed animals (3 hours) for comparisons to previoulsy collected starved data to see what acute effects nutritional flux would have on either O-GlcNAc chromatin marks or RNA Pol II distributions. L1 larvae were synchronized by starvation for two days after hatching in sterile buffer and then plated on NGM plates seeded with E. coli (OP50) and allowed to feed for 3 hours prior to collection. Wild type and mutant animals were collected, washed, and chromatin isolated for immunoprecipitation with a variety of antibodies that recognize either O-GlcNAcylated substrates or RNA Pol II.