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. Author manuscript; available in PMC: 2010 Mar 10.
Published in final edited form as: Nature. 2009 Aug 23;461(7261):225–229. doi: 10.1038/nature08265

Figure 1. Decapped mRNA is associated with polyribosomes.

Figure 1

(a) Primer extension analysis on endogenous PGK1, CYH2, and ADH1 mRNA was performed on RNA isolated from sucrose gradient fractions of an xrn1Δ cell lysate. RNP, 80S, and polyribosomes are indicated above fraction numbers. FL, full length mRNA; - cap, decapped mRNA. Primer extension analyses on total RNA (15 μg) from WT, dcp2Δ, and xrn1Δ cells are shown on left side of each panel to indicate −cap mRNA is observed only in xrn1Δ cells. (b) Representation of PGK1 reporter, PGK1 reporter with a PTC (PGK1short), and PGK1 reporter with a stem-loop in its 5’ UTR (SL-PGK1). (c) Primer extension on RNA from sucrose gradient fractions from lysates of upf1Δ/xrn1Δ cells expressing PGK1 reporter or PGK1short reporter, and from xrn1Δ cells expressing SL-PGK1 or PGK1 reporter. In the bottom panel, lysates from xrn1Δ cells expressing the PGK1 reporter were incubated in presence of 50 mM EDTA prior to loading on sucrose gradients. (d) Quantification of – cap mRNAs as a percentage of total reverse transcription product in RNP and polyribosome fractions.