Ribosomal protein eL42 contributes to the catalytic activity of the yeast ribosome at the elongation step of translation. / Hountondji, Codjo; Créchet, Jean Bernard; Tanaka, Mayo et al.
In: Biochimie, Vol. 158, 03.2019, p. 20-33.Research output: Contribution to journal › Article › peer-review
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TY - JOUR
T1 - Ribosomal protein eL42 contributes to the catalytic activity of the yeast ribosome at the elongation step of translation
AU - Hountondji, Codjo
AU - Créchet, Jean Bernard
AU - Tanaka, Mayo
AU - Suzuki, Mieko
AU - Nakayama, Jun ichi
AU - Aguida, Blanche
AU - Bulygin, Konstantin
AU - Cognet, Jean
AU - Karpova, Galina
AU - Baouz, Soria
N1 - Copyright © 2018 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.
PY - 2019/3
Y1 - 2019/3
N2 - 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.
AB - 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.
KW - eL42 protein from human or Schizosaccharomyces pombe
KW - Eukaryal 80S ribosomes
KW - Lys-55 of S. pombe eL42
KW - Mechanism of peptidyl transfer and peptidyl-tRNA hydrolysis by eukaryotic 80S ribosomes
KW - the GGQ motif of eL42
KW - The peptidyl transferase center of 80S ribosomes from eukaryotes
KW - Schizosaccharomyces/chemistry
KW - Mutation, Missense
KW - Peptide Chain Elongation, Translational/physiology
KW - Schizosaccharomyces pombe Proteins/chemistry
KW - Ribosomes/chemistry
KW - Ribosomal Proteins/chemistry
KW - Catalysis
KW - Amino Acid Substitution
KW - CYCLOHEXIMIDE RESISTANCE
KW - GGQ MOTIF
KW - ESCHERICHIA-COLI
KW - PEPTIDE-BOND FORMATION
KW - TRANSFER RNA-SYNTHETASE
KW - RELEASE FACTOR ERF1
KW - TRANSFERASE CENTER
KW - MESSENGER-RNA
KW - STRUCTURAL BASIS
KW - BINDING-SITE
UR - http://www.scopus.com/inward/record.url?scp=85058559263&partnerID=8YFLogxK
U2 - 10.1016/j.biochi.2018.12.005
DO - 10.1016/j.biochi.2018.12.005
M3 - Article
C2 - 30550856
AN - SCOPUS:85058559263
VL - 158
SP - 20
EP - 33
JO - Biochimie
JF - Biochimie
SN - 0300-9084
ER -
ID: 22848698