Loss of Elongator- and KEOPS-Dependent tRNA Modifications Leads to Severe Growth Phenotypes and Protein Aggregation in Yeast

Biomolecules. 2020 Feb 18;10(2):322. doi: 10.3390/biom10020322.

Abstract

Modifications found in the Anticodon Stem Loop (ASL) of tRNAs play important roles in regulating translational speed and accuracy. Threonylcarbamoyl adenosine (t6A37) and 5-methoxycarbonyl methyl-2-thiouridine (mcm5s2U34) are critical ASL modifications that have been linked to several human diseases. The model yeast Saccharomyces cerevisiae is viable despite the absence of both modifications, growth is however greatly impaired. The major observed consequence is a subsequent increase in protein aggregates and aberrant morphology. Proteomic analysis of the t6A-deficient strain (sua5 mutant) revealed a global mistranslation leading to protein aggregation without regard to physicochemical properties or t6A-dependent or biased codon usage in parent genes. However, loss of sua5 led to increased expression of soluble proteins for mitochondrial function, protein quality processing/trafficking, oxidative stress response, and energy homeostasis. These results point to a global function for t6A in protein homeostasis very similar to mcm5/s2U modifications.

Keywords: protein aggregation; tRNA modification.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Anticodon / genetics
  • Anticodon / metabolism
  • DNA-Binding Proteins / genetics*
  • DNA-Binding Proteins / metabolism
  • Histone Acetyltransferases / genetics*
  • Histone Acetyltransferases / metabolism
  • Nucleic Acid Conformation
  • Phenotype
  • Protein Aggregates / physiology
  • Protein Biosynthesis / genetics
  • Protein Biosynthesis / physiology
  • Proteins / genetics
  • Proteomics / methods
  • RNA, Transfer / genetics
  • RNA, Transfer / metabolism*
  • Saccharomyces cerevisiae / growth & development
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / genetics*
  • Saccharomyces cerevisiae Proteins / metabolism
  • Thermodynamics
  • Thiouridine / analogs & derivatives
  • Thiouridine / chemistry

Substances

  • 5-carbomethoxymethyl-2-thiouridine
  • Anticodon
  • DNA-Binding Proteins
  • Protein Aggregates
  • Proteins
  • SUA5 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Thiouridine
  • RNA, Transfer
  • Elp3 protein, S cerevisiae
  • Histone Acetyltransferases