Accurate and fast graph-based pangenome annotation and clustering with ggCaller

Bacterial genomes differ in both gene content and sequence mutations, which can cause important clinical phenotypic differences such as vaccine escape or antimicrobial resistance. To identify and quantify important variants, all genes within a population must be predicted, functionally annotated and clustered, representing the ‘pangenome’. Despite the volume of genome data available, gene prediction and annotation are currently conducted in isolation on individual genomes, which is computationally inefficient and frequently inconsistent across genomes. Here, we introduce the open-source software graph-gene-caller (ggCaller; https://github.com/samhorsfield96/ggCaller ). ggCaller combines gene prediction, functional annotation and clustering into a single step using population-wide de Bruijn Graphs, removing redundancy in gene annotation, and resulting in more accurate gene predictions and orthologue clustering. We applied ggCaller to simulated and real-world bacterial genome datasets, comparing it to current state-of-the-art tools. ggCaller is ~50x faster with equivalent or greater accuracy, particularly in datasets with complex sources of error, such as assembly contamination or fragmentation. ggCaller is also an important extension to bacterial genome-wide association studies, enabling querying of annotated graphs for functional analyses. We highlight this application by functionally annotating DNA sequences with significant associations to tetracycline and macrolide resistance in Streptococcus pneumoniae , identifying key resistance determinants that were missed when using only a single reference genome. ggCaller is a novel bacterial genome analysis tool with applications in bacterial epidemiology and evolutionary study.