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. 2019 Dec 13;17(12):e3000329. doi: 10.1371/journal.pbio.3000329

Fig 1. Nob1 prevents entry of pre-40S subunits into the polysomes.

Fig 1

(A) Shown are 10%–50% sucrose gradients from lysates of cells depleted of Nob1 by growth in glucose for 12 h. Shown below the absorbance profile at 254 nm are northern blots of pre-18S rRNA (20S), mature 18S, and 25S rRNAs and western blots probing for Pno1. Arrowhead notes the upper band corresponding to Pno1. (B) The effects from depletion of Nob1 on the fidelity of start codon recognition, decoding, stop codon recognition, and FS (−1 and +1) were assayed using dual-luciferase reporters. Shown are the relative error rates of the glucose-depleted samples relative to replete samples. Data are the averages of 10−27 biological replicates, and error bars indicate the SEM. *p < 0.05, **p < 0.01, ***p < 0.001 by unpaired t test. (C) Changes in doubling time in cells replete (Nob1) or depleted (ΔNob1) for Nob1 after exposure to high salt (1 M NaCl) or caffeine (10 mM). Values were compared to no-stress conditions (fold change = 1). Data are the averages of six (caffeine) or four to five (high salt) biological replicates, and error bars indicate SEM. **p < 0.01 by unpaired t test. Numerical data are listed in S1 Data. (D, E) Sucrose gradients of wild-type BY4741 cells overexpressing Nob1 (D) or Nob1-D15N (E) under the galactose-inducible, glucose-repressible Gal promoter in galactose for 12 h. Western blots probed for Nob1. Arrowhead notes the upper band corresponding to Nob1. (F) Sucrose gradients of WT BY4741 cells overexpressing Nob1 under the constitutive Tef2 promoter grown in glucose. See also S1 Fig. FS, frameshifting; Gal, galactose; WT, wild-type.