Fig. 2: Large-scale pooled phenotypic screens enable identification of processes underlying the P. falciparum heat-shock response.

a HS-Sensitive mutations identified in pooled screens of the 1K-library of pB-mutants (Supplementary Data File 2). The uncloned, large mixed-population pools comprising the 1K-library (n = 12) were parallelly screened in both ideal growth conditions and under HS. Mutants were assigned phenotypes as per methods established in the pilot-library screens (Fig. 1 and Supplementary Data File 2). Mutants are ranked by fold-change from HS-Sensitive (red; n = 149, FC-HS < 0.5 and PFS_HS < 0.25) to HS-Tolerant (green; n = 139, FC-HS > 1.5). Mean mutant fold-change in ideal growth (FC-Growth) is superimposed as a bar plot (gray, FC-Growth <1.0; blue, FC-Growth > 1.0). Mutants performing poorly in both screens (yellow; n = 91, FC-HS < 0.5, PFS_HS > 0.25) were classified as HS- and Growth-Sensitive and were not considered further (Supplementary Fig. 9). Mutations neither HS-Sensitive nor HS-Tolerant were classified as HS-Neutral (taupe, n = 543). Internal redundancy within the 1K- and pilot clone libraries allowed assessment of correlation between identical or same-gene mutants across pools. Phenotypes were highly reproducible. See Supplementary Figs. 6a–c, 7a–b, 8a, b. b Functional enrichment of GO terms for HS-Sensitive or Growth-Sensitive pB-mutants vs all other mutants in the 1K-library. HS-Sensitive mutants were enriched in terms associated with HS-response such as protein folding, response to DNA-damage, DNA-repair, and regulation of vesicle-mediated transport. Growth-Sensitive mutants tended to be enriched for more general categories broadly important for survival in all conditions, such as translation- or mRNA-metabolism-related terms. Circles represent the GO category, circle color represents ontology, and circle size represents the number of significant genes annotated to that category. Significant terms (two-tailed Fisher/elim-hybrid test p-value ≤ 0.05) fall within the light-green box.