Ribosomal protein eL42 contributes to the catalytic activity of the yeast ribosome at the elongation step of translation

Codjo Hountondji, Jean Bernard Créchet, Mayo Tanaka, Mieko Suzuki, Jun ichi Nakayama, Blanche Aguida, Konstantin Bulygin, Jean Cognet, Galina Karpova, Soria Baouz

Research output: Contribution to journalArticlepeer-review

3 Citations (Scopus)

Abstract

The GGQ minidomain of the ribosomal protein eL42 was previously shown to contact the CCA-arm of P-site bound tRNA in human ribosome, indicating a possible involvement of the protein in the catalytic activity. Here, using Schizosaccharomyces pombe (S. pombe) cells, we demonstrate that the GGQ minidomain and neighboring region of eL42 is critical for the ribosomal function. Mutant eL42 proteins containing amino acid substitutions within or adjacent to the GGQ minidomain failed to complement the function of wild-type eL42, and expression of the mutant eL42 proteins led to severe growth defects. These results suggest that the mutations in eL42 interfere with the ribosomal function in vivo. Furthermore, we show that some of the mutations associated with the conserved GGQ region lead to reduced activities in the poly(Phe) synthesis and/or in the peptidyl transferase reaction with respect to puromycin, as compared with those of the wild-type ribosomes. A pK value of 6.95 was measured for the side chain of Lys-55/Arg-55, which is considerably less than that of a Lys or Arg residue. Altogether, our findings suggest that eL42 contributes to the 80S ribosome's peptidyl transferase activity by promoting the course of the elongation cycle.

Original languageEnglish
Pages (from-to)20-33
Number of pages14
JournalBiochimie
Volume158
DOIs
Publication statusPublished - Mar 2019
Externally publishedYes

Keywords

  • eL42 protein from human or Schizosaccharomyces pombe
  • Eukaryal 80S ribosomes
  • Lys-55 of S. pombe eL42
  • Mechanism of peptidyl transfer and peptidyl-tRNA hydrolysis by eukaryotic 80S ribosomes
  • the GGQ motif of eL42
  • The peptidyl transferase center of 80S ribosomes from eukaryotes
  • Schizosaccharomyces/chemistry
  • Mutation, Missense
  • Peptide Chain Elongation, Translational/physiology
  • Schizosaccharomyces pombe Proteins/chemistry
  • Ribosomes/chemistry
  • Ribosomal Proteins/chemistry
  • Catalysis
  • Amino Acid Substitution
  • CYCLOHEXIMIDE RESISTANCE
  • GGQ MOTIF
  • ESCHERICHIA-COLI
  • PEPTIDE-BOND FORMATION
  • TRANSFER RNA-SYNTHETASE
  • RELEASE FACTOR ERF1
  • TRANSFERASE CENTER
  • MESSENGER-RNA
  • STRUCTURAL BASIS
  • BINDING-SITE

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