TY - JOUR T1 - Transposon insertional mutagenesis in <em>Saccharomyces uvarum</em> reveals <em>trans</em>-acting effects influencing species-dependent essential genes JF - bioRxiv DO - 10.1101/218305 SP - 218305 AU - Monica R. Sanchez AU - Celia Payen AU - Frances Cheong AU - Blake T. Hovde AU - Sarah Bissonnette AU - Adam P. Arkin AU - Jeffrey M. Skerker AU - Rachel B. Brem AU - Amy A. Caudy AU - Maitreya J. Dunham Y1 - 2018/01/01 UR - http://biorxiv.org/content/early/2018/11/07/218305.abstract N2 - To understand how complex genetic networks perform and regulate diverse cellular processes, the function of each individual component must be defined. Comprehensive phenotypic studies of mutant alleles have been successful in model organisms in determining what processes depend on the normal function of a gene. These results are often ported to newly sequenced genomes by using sequence homology. However, sequence similarity does not always mean identical function or phenotype, suggesting that new methods are required to functionally annotate newly sequenced species. We have implemented comparative analysis by high-throughput experimental testing of gene dispensability in Saccharomyces uvarum, a sister species of S. cerevisiae. We created haploid and heterozygous diploid Tn7 insertional mutagenesis libraries in S. uvarum to identify species dependent essential genes, with the goal of detecting genes with divergent functions and/or different genetic interactions. Comprehensive gene dispensability comparisons with S. cerevisiae predicted diverged dispensability at 12% of conserved orthologs, and validation experiments confirmed 22 differentially essential genes. Surprisingly, despite their differences in essentiality, these genes were capable of cross-species complementation, demonstrating that trans-acting factors that are background-dependent contribute to differential gene essentiality. This study demonstrates that direct experimental testing of gene disruption phenotypes across species can inform comparative genomic analyses and improve gene annotation. Our method can be widely applied in microorganisms to further our understanding of genome evolution. ER -