Genetic Interaction of tRNA-Dependent Mistranslation with Fused in Sarcoma Protein Aggregates

High-fidelity protein synthesis requires properly aminoacylated transfer RNAs (tRNAs), yet diverse cell types, from bacteria to humans, show a surprising ability to tolerate errors in translation resulting from mutations in tRNAs, aminoacyl-tRNA synthetases, and other components of protein synthesis. Recently, we characterized a tRNA(AGA)(Ser) G35A mutant (tRNA(AAA)(Ser)) that occurs in 2% of the human population. The mutant tRNA decodes phenylalanine codons with serine, inhibits protein synthesis, and is defective in protein and aggregate degradation. Here, we used cell culture models to test our hypothesis that tRNA-dependent mistranslation will exacerbate toxicity caused by amyotrophic lateral sclerosis (ALS)-associated protein aggregation. Relative to wild-type tRNA, we found cells expressing tRNA(AAA)(Ser) showed slower but effective aggregation of the fused in sarcoma (FUS) protein. Despite reduced levels in mistranslating cells, wild-type FUS aggregates showed similar toxicity in mistranslating cells and normal cells. The aggregation kinetics of the ALS-causative FUS R521C variant were distinct and more toxic in mistranslating cells, where rapid FUS aggregation caused cells to rupture. We observed synthetic toxicity in neuroblastoma cells co-expressing the mistranslating tRNA mutant and the ALS-causative FUS R521C variant. Our data demonstrate that a naturally occurring human tRNA variant enhances cellular toxicity associated with a known causative allele for neurodegenerative disease.

Automated summary

High-fidelity protein synthesis relies on properly aminoacylated tRNAs, but cells have the ability to tolerate errors in translation due to mutations in tRNAs and other components. A mutant tRNA variant, tRNA(AAA)(Ser), was found in 2% of the human population and showed defective protein synthesis and degradation. In this study, we demonstrated that this tRNA variant enhanced the toxicity of ALS-associated protein aggregation, particularly with the ALS-causative FUS R521C variant, leading to cell rupture and synthetic toxicity in neuroblastoma cells.