Expression from C. elegans L1 animals. Caenorhabditis elegans

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. Here we compare gene expression in wild type and O-GlcNAc mutants (ogt-1 and oga-1) in synchronized, fed L1 animals. Whole genome transcriptional profiling of the O-GlcNAc cycling mutants confirmed dramatic deregulation of genes in these key pathways. As predicted, the O-GlcNAc cycling mutants show phenotypically altered lifespan and susceptibility to UV stress. Overall design: L1 larvae were synchronized by starvation for two days after hatching in sterile buffer and then plated on NGM with E. coli (OP50) and allowed to feed for 3 hours at room temperature. Wild type and mutant animals were collected and total RNA isolated for gene expression analysis.