Fluoropyrimidines trigger decay of hypomodified tRNA in yeast

Nucleic Acids Res. 2024 Jun 10;52(10):5841-5851. doi: 10.1093/nar/gkae341.

Abstract

Therapeutic fluoropyrimidines 5-fluorouracil (5-FU) and 5-fluorocytosine (5-FC) are in long use for treatment of human cancers and severe invasive fungal infections, respectively. 5-Fluorouridine triphosphate represents a bioactive metabolite of both drugs and is incorporated into target cells' RNA. Here we use the model fungus Saccharomyces cerevisiae to define fluorinated tRNA as a key mediator of 5-FU and 5-FC cytotoxicity when specific tRNA methylations are absent. tRNA methylation deficiency caused by loss of Trm4 and Trm8 was previously shown to trigger an RNA quality control mechanism resulting in partial destabilization of hypomodified tRNAValAAC. We demonstrate that, following incorporation into tRNA, fluoropyrimidines strongly enhance degradation of yeast tRNAValAAC lacking Trm4 and Trm8 dependent methylations. At elevated temperature, such effect occurs already in absence of Trm8 alone. Genetic approaches and quantification of tRNA modification levels reveal that enhanced fluoropyrimidine cytotoxicity results from additional, drug induced uridine modification loss and activation of tRNAValAAC decay involving the exonuclease Xrn1. These results suggest that inhibition of tRNA methylation may be exploited to boost therapeutic efficiency of 5-FU and 5-FC.

MeSH terms

  • Exoribonucleases / genetics
  • Exoribonucleases / metabolism
  • Flucytosine* / pharmacology
  • Fluorouracil* / pharmacology
  • Methylation
  • RNA Stability / drug effects
  • RNA, Transfer* / metabolism
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism
  • Saccharomyces cerevisiae* / drug effects
  • Saccharomyces cerevisiae* / genetics
  • Saccharomyces cerevisiae* / metabolism
  • Uridine / metabolism
  • tRNA Methyltransferases / genetics
  • tRNA Methyltransferases / metabolism

Substances

  • Exoribonucleases
  • Flucytosine
  • Fluorouracil
  • RNA, Transfer
  • Saccharomyces cerevisiae Proteins
  • tRNA Methyltransferases
  • Uridine
  • XRN1 protein, S cerevisiae
  • NCL1 protein, S cerevisiae
  • tRNA (guanine-N7-)-methyltransferase