Adaptation to overflow metabolism by mutations that impair tRNA modification in experimentally evolved bacteria

• Published in: bioRxiv
• Main Authors: Muraski, Marc, Nilsson, Emil M, Fritz, Melissa J, Richardson, Anthony R, Alexander, Rebecca M, Cooper, Vaughn S
• Format: Paper
• Language: English
• Published: Cold Spring Harbor Cold Spring Harbor Laboratory Press 30.10.2022; Cold Spring Harbor Laboratory
• Edition: 1.1
• Subjects: Galactose; Homeostasis; Lysine; Metabolism; Microbiology; Mutants; Mutation; Oxidative metabolism; Positive selection; Reproductive fitness; tRNA; tRNA Ala
• ISSN: 2692-8205; 2692-8205
• DOI: 10.1101/2022.10.28.514323

When microbes grow in foreign nutritional environments, selection may enrich mutations in unexpected pathways connecting growth and homeostasis. An evolution experiment designed to identify beneficial mutations in Burkholderia cenocepacia captured six independent nonsynonymous substitutions in the essential gene tilS, which modifies tRNAIle2 by adding a lysine to the anticodon for faithful AUA recognition. Further, five additional mutants acquired mutations in tRNAIle2, which strongly suggests that disrupting the TilS:tRNAIle2 interaction was subject to strong positive selection. Mutated TilS incurred greatly reduced enzymatic function but retained capacity for tRNAIle2 binding. Yet both mutant sets outcompeted wild-type by decreasing lag phase duration by ~3.5 hours. We hypothesized that lysine demand could underlie fitness in the experimental conditions. As predicted, supplemental lysine complemented the ancestral fitness deficit, but so did additions of several other amino acids. Mutant fitness advantages were also specific to rapid growth on galactose using oxidative overflow metabolism that generates redox imbalance, not resources favoring more balanced metabolism. Remarkably, 13 tilS mutations also evolved in the Long-Term Evolution Experiment with E. coli, including four fixed mutations. These results suggest that TilS or unknown binding partners contribute to improved growth under conditions of rapid sugar oxidation at the predicted expense of translational accuracy. Competing Interest Statement The authors have declared no competing interest.