Fig. 1. Genome-wide mapping of genetic interactions with human NatC.

a Schematic of genome-wide CRISPR screens to identify genetic interactions (GI) with NatC. HAP1 WT, NAA30-KO, NAA35-KO, and NAA38-KO cells were transduced with a pooled genome-wide CRISPR knockout library (TKOv3) and selected for viral integration. gRNA regions were PCR-amplified from genomic DNA extracted from cells collected at the start (T0) and endpoint of the screen (T14-18). gRNA abundance was determined by next-generation sequencing (NGS). b, c Reproducibility of NAA35 qGI scores. b qGI scores were determined by comparing the log₂-fold change (LFC) for every gene represented in the TKOv3 library in NAA35-KO cell line with those observed in a panel of WT control screens. Pearson correlation coefficient (r) was calculated using all qGI scores (r in black, calculated from all data points) or using a stringent cut-off for the GIs (|qGI|>0.3, FDR < 0.10) in both screens (r and datapoints marked in purple). c The Pearson correlation coefficients of the qGI scores (two replicated screens) was adjusted to the similarity of a NAA35-KO screen to a panel of HAP1-KO screens. The resulting Within vs Between replicate Correlation (WBC) score provides a confidence of reproducibility interpreted as a z-score. d Negative and positive GIs of NAA35. Scatterplot showing the fitness effect (LFC) of 486 genes in NAA35-KO versus WT cells, showing a significant GI in at least two NAA35 screens (|qGI|>0.3, FDR < 0.10). Negative (blue) and positive (yellow) NAA35 GIs are shown. Darker color indicates interactions that were called in all three replicate screens. Node size corresponds to strength of the mean absolute GI score (three independent screens). Volcano plots displaying qGI scores and associated significance (log₁₀ values) for genes targeted by the TKOv3 library in (e) NAA35-KO, (f) NAA30-KO and (g) NAA38-KO screens. Negative (blue) and positive (yellow) GIs are shown. h–j Negative GIs of NatC indicate a role in Golgi transport. Pathway enrichment analysis of genes exhibiting a negative GI with (h) NAA35, (i) NAA30 or (j) NAA38 (identified in at least two NAA35 screens; |qGI|>0.3, FDR < 0.1). Benjamini–Hochberg adjusted p-values for each gene ontology term is indicated by gray gradient.