Fig. 5: Quantifying fitness of mutants that incorporate WT phosphosites.

a Schematic showing the design of the WT/A plasmid library. S/T-P sites within the TAD were either left intact or mutated to A-P in all possible combinations (256). b, c Box and whisker plots showing the selection coefficients of groups of mutants with the indicated number of WT (W) S/T-P sites (b), or with specific substitutions at the indicated positions (c). Color gradient from blue (hcm1-8A) to red (HCM1) corresponds to increasing numbers of WT sites (b). Blue represents all mutants with an A-P mutation at the indicated site, gray represents all mutants with wild-type sequences at the indicated site (c). The black center line indicates the median, boxes indicate the 25th−75th percentiles, black lines represent 1.5 IQR of the 25th and 75th percentile, black circles represent outliers. Shown is data from n = 3 biological replicates. See Supplementary Fig. 3c for correlations between replicates and Supplementary Fig. 6a for heatmap of screen results. d Schematic showing the design of the WT/E plasmid library. S/T-P sites in the TAD were either left intact or mutated to E-E in all possible combinations (256). e, f Box and whisker plots showing the selection coefficients of groups of mutants with the indicated number of phosphomimetic mutations (e), or with specific substitutions at the indicated positions (f). Color gradient from blue (HCM1) to red (hcm1-8E) corresponds to increasing numbers of phosphomimetic mutations (e). Gray represents all mutants with WT sequence at the indicated site, red represents all mutants that have an E-E mutation at the indicated site (f). The black center line indicates the median, boxes indicate the 25th−75th percentiles, black lines represent 1.5 IQR of the 25th and 75th percentile, black circles represent outliers. Shown is data from n = 3 biological replicates. See Supplementary Fig. 3d for correlations between replicates and Supplementary Fig. 6b, c for heatmaps of screen results.