Fig. 3: Osmotic stress-induced changes in metabolism changes the spectrum of mutants buffered. | Nature Communications

Fig. 3: Osmotic stress-induced changes in metabolism changes the spectrum of mutants buffered.

From: Distinct metabolic states of a cell guide alternate fates of mutational buffering through altered proteostasis

Fig. 3

a Gene Ontology (GO) classes upregulated in WG350(S) transcriptome with respect to CSH4(S) shown as fold enrichment on the left-axis and Benjamini-Hochberg FDR corrected p-values obtained using DAVID on the right axis from two biological replicates. b Comparison of metabolites in WG350 and CSH4 during growth in media containing 350 mM NaCl using untargeted metabolomics. Colored circles represent significantly different metabolites obtained using paired 2-sided Student’s t-test. c Comparison of metabolites within strain in presence and absence of NaCl. Colored circles represent significantly different metabolites obtained using paired 2-sided Student’s t-test. d Mean of normalized read counts of Gm-R GG-mutant library at comparative selection pressure of Gentamicin (Gm) and 350 mM NaCl in growth media from two biological replicates. Pink shaded area marks 99% confidence interval. e Heatmap representing activity of small-molecule-dependent Gm-R mutants in CSH4(S) and WG350(S) at different concentrations of Gentamicin (Gm) (top panel). Bottom panel shows mutant MIC normalized with respect to Wt Gm-R in the respective strains. Mean with error bars representing standard deviation from four biological replicates is plotted (2-sided Student’s t-test p-value > 0.05). f The scatter plot for mCherry Vs GFP/mCherry fluorescence of pool of GFP mutants with compromised fluorescence (Pc) in WG350(S) and CSH4(S). Blue box indicates clones buffered in WG350(S). g Histogram of GFP/mCherry fluorescence of Wt, C1, C3, and C6 GFP in WG350 and CSH4 in presence and absence of osmotic stress. Cyan arrow indicates increase in GFP fluorescence upon addition of 350 mM NaCl to CSH4 strain, orange arrow indicates the same for WG350. h Conservation score (calculated using Consurf) for each residue. Dots indicate residues that are mutated in the clones studied. i Fractional ASA (calculated using VADAR) of the residues mutated in the clones is plotted against ΔΔG of the residues obtained from 1GFL. Error bars represent standard deviation from three replicates (calculated using FoldX). For b and c: p-value < 0.05, five biological replicates per sample. Also see Supplementary Fig. 2 (Source data are provided as source data file Fig. 3).

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