Analyzing P-bodies in Saccharomyces cerevisiae

Abstract

Cytoplasmic processing bodies, or P-bodies, are RNA-protein granules found in eukaryotic cells. P-bodies contain non-translating mRNAs and proteins involved in mRNA degradation and translational repression. P-bodies, and the mRNPs within them, have been implicated in mRNA storage, mRNA degradation, and translational repression. The analysis of mRNA turnover often involves the analysis of P-bodies. In this chapter, we describe methods to analyze P-bodies in the budding yeast, Saccharomyces cerevisiae, including procedures to determine whether a protein or mRNA can accumulate in P-bodies, whether an environmental perturbation or mutation affects P-body size and number, and methods to quantify P-bodies.

Figure 1. P-bodies under different growth and induction conditions

The P-body marker used is the genomically integrated DCP2-GFP (yRP2162). The cells were grown in YEPD to (A) mid log growth phase, then treated with (B) glucose deprivation for 10’ in complete media or (C) hyperosmotic (1 M KCl) stress for 15’ in complete minimal media with glucose as described in the text.