Cell Reports
Volume 7, Issue 2, 24 April 2014, Pages 339-347
Journal home page for Cell Reports

Report
Structure, Mechanism, and Specificity of a Eukaryal tRNA Restriction Enzyme Involved in Self-Nonself Discrimination

https://doi.org/10.1016/j.celrep.2014.03.034Get rights and content
Under a Creative Commons license
open access

Highlights

  • Microbes secrete tRNA anticodon nuclease toxins to thwart nonself species

  • Fungal ribotoxin PaT incises tRNAGln anticodon stem-loop 3′ of the wobble uridine

  • The crystal structure of PaT reveals a distinctive fold and active site

  • Toxin immunity is conferred by ImmPaT, a tRNase inhibitor

Summary

tRNA restriction by anticodon nucleases underlies cellular stress responses and self-nonself discrimination in a wide range of taxa. Anticodon breakage inhibits protein synthesis, which, in turn, results in growth arrest or cell death. The eukaryal ribotoxin PaT secreted by Pichia acaciae inhibits growth of Saccharomyces cerevisiae via cleavage of tRNAGln(UUG). We find that recombinant PaT incises a synthetic tRNAGln(UUG) stem-loop RNA by transesterification at a single site 3′ of the wobble uridine, yielding 2′,3′-cyclic phosphate and 5′-OH ends. Incision is suppressed by replacement of the wobble nucleobase with adenine or guanine. The crystal structure of PaT reveals a distinctive fold and active site, essential components of which are demonstrated by mutagenesis. Pichia acaciae evades self-toxicity via a distinctive intracellular immunity protein, ImmPaT, which binds PaT and blocks nuclease activity. Our results highlight the evolutionary diversity of tRNA restriction and immunity systems.

Cited by (0)

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/).