Modification of tRNA(Lys) UUU by elongator is essential for efficient translation of stress mRNAs

PLoS Genet. 2013;9(7):e1003647. doi: 10.1371/journal.pgen.1003647. Epub 2013 Jul 18.

Abstract

The Elongator complex, including the histone acetyl transferase Sin3/Elp3, was isolated as an RNA polymerase II-interacting complex, and cells deficient in Elongator subunits display transcriptional defects. However, it has also been shown that Elongator mediates the modification of some tRNAs, modulating translation efficiency. We show here that the fission yeast Sin3/Elp3 is important for oxidative stress survival. The stress transcriptional program, governed by the Sty1-Atf1-Pcr1 pathway, is affected in mutant cells, but not severely. On the contrary, cells lacking Sin3/Elp3 cannot modify the uridine wobble nucleoside of certain tRNAs, and other tRNA modifying activities such as Ctu1-Ctu2 are also essential for normal tolerance to H2O2. In particular, a plasmid over-expressing the tRNA(Lys) UUU complements the stress-related phenotypes of Sin3/Elp3 mutant cells. We have determined that the main H2O2-dependent genes, including those coding for the transcription factors Atf1 and Pcr1, are highly expressed mRNAs containing a biased number of lysine-coding codons AAA versus AAG. Thus, their mRNAs are poorly translated after stress in cells lacking Sin3/Elp3 or Ctu2, whereas a mutated atf1 transcript with AAA-to-AAG lysine codons is efficiently translated in all strain backgrounds. Our study demonstrates that the lack of a functional Elongator complex results in stress phenotypes due to its contribution to tRNA modification and subsequent translation inefficiency of certain stress-induced, highly expressed mRNAs. These results suggest that the transcriptional defects of these strain backgrounds may be a secondary consequence of the deficient expression of a transcription factor, Atf1-Pcr1, and other components of the transcriptional machinery.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatin / drug effects
  • Chromatin / genetics
  • Chromatin / metabolism
  • Histone Acetyltransferases / genetics
  • Histone Acetyltransferases / metabolism
  • Hydrogen Peroxide / pharmacology
  • Lysine / metabolism
  • Oxidative Stress / genetics*
  • Peptide Chain Elongation, Translational
  • Protein Biosynthesis / drug effects
  • RNA Polymerase II / genetics
  • RNA Polymerase II / metabolism
  • RNA, Messenger / genetics*
  • RNA, Messenger / metabolism
  • RNA, Transfer, Lys / genetics
  • RNA, Transfer, Lys / metabolism*
  • Schizosaccharomyces / genetics*
  • Schizosaccharomyces pombe Proteins / genetics
  • Schizosaccharomyces pombe Proteins / metabolism
  • Uridine / genetics

Substances

  • Chromatin
  • RNA, Messenger
  • RNA, Transfer, Lys
  • Schizosaccharomyces pombe Proteins
  • Hydrogen Peroxide
  • Elp3 protein, S pombe
  • Histone Acetyltransferases
  • RNA Polymerase II
  • Lysine
  • Uridine

Grants and funding

This work was supported by the Spanish Ministry of Science and Innovation (BFU2009-06933 and BFU2012-32045), PLAN E and FEDER, by the Spanish program Consolider-Ingenio 2010 Grant CSD 2007-0020, and by SGR2009-196 from Generalitat de Catalunya (Spain) to EH. EH and JA are recipients of ICREA Academia Awards (Generalitat de Catalunya). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.