Ybr094wp (Pby1p) localizes in P-bodies and plays no obvious role in 5′–3′ mRNA decay, NMD, or decay of EDC1 mRNA. (A) Cells encoding a chromosomal copy of YBR094W-GFP were transformed with DCP2-RFP encoded on a plasmid, and cultures were treated with medium either containing or lacking glucose for 10 min, at which time DAPI was added directly to the culture medium. Cells were then imaged by fluorescence microscopy. Half-lives of MFA2pG mRNA (B) and PGK1pG mRNA (C) were determined in cultures of wild-type or pby1Δ cells by transcriptional shut-off by glucose repression. RNA was isolated at various time points after repression (indicated in the figure) and analyzed by Northern blot. Steady-state levels of CYH2 pre-mRNA and EDC1 mRNA were determined in cultures of wild-type, pby1Δ, and xrn1Δ cells by RNA isolation followed by Northern blot analysis (D).
