The human large subunit ribosomal protein L36A-like contacts the CCA end of P-site bound tRNA

Baouz, Soria; Woisard, Anne; Sinapah, Sylvie; Caër, Jean-Pierre Le; Argentini, Manuela; Bulygin, K. N.; Aguié, Gustave; Hountondji, Codjo

doi:10.1016/j.biochi.2009.07.013

Cited by 30 publications

(42 citation statements)

References 30 publications

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“…Together, these observations support the possibility that the loop extension of Rpl42 and the methylation at lysine 55 play a critical role in an interaction with the deacylated tRNA positioned at the E-site. Intriguingly, human L36a-like, which is closely related to Rpl36a/Rpl42, has been demonstrated to make contact with the CCA end of P-site-bound tRNA (55). Therefore, it is also possible that Rpl42 plays a role in the recognition of tRNA positioned at the P-site.…”

Section: Discussionmentioning

confidence: 99%

Methylation of Ribosomal Protein L42 Regulates Ribosomal Function and Stress-adapted Cell Growth

Shirai

Sadaie

Shinmyozu³

et al. 2010

Journal of Biological Chemistry

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Lysine methylation is one of the most common protein modifications. Although lysine methylation of histones has been extensively studied and linked to gene regulation, that of nonhistone proteins remains incompletely understood. Here, we show a novel regulatory role of ribosomal protein methylation. Using an in vitro methyltransferase assay, we found that Schizosaccharomyces pombe Set13, a SET domain protein encoded by SPAC688.14, specifically methylates lysine 55 of ribosomal protein L42 (Rpl42). Mass spectrometric analysis revealed that endogenous Rpl42 is monomethylated at lysine 55 in wild-type S. pombe cells and that the methylation is lost in ⌬set13 mutant cells. ⌬set13 and Rpl42 methylation-deficient mutant S. pombe cells showed higher cycloheximide sensitivity and defects in stress-responsive growth control compared with wild type. Genetic analyses suggested that the abnormal growth phenotype was distinct from the conserved stress-responsive pathway that modulates translation initiation. Furthermore, the Rpl42 methylation-deficient mutant cells showed a reduced ability to survive after entering stationary phase. These results suggest that Rpl42 methylation plays direct roles in ribosomal function and cell proliferation control independently of the general stress-response pathway.Post-translational modifications regulate protein structure and function, and one of the most common is lysine methylation (1). Although lysine methylation does not change the overall charge of the side chain, it can influence the activity of protein by providing a novel interface for its interactions with other molecules. Most protein lysine methylations, with a few exceptions, are catalyzed by the SET domain-containing protein (SET protein) family (2). The SET domain was originally identified in three Drosophila proteins, Su(var)3-9, Enhancer of zest, and Trithorax, and was later demonstrated to be the responsible domain for histone-lysine methyltransferase function (3). The characterization of these enzymes has revealed the roles of histone lysine methylation in many different biological processes, including higher order chromatin assembly and transcriptional regulation (4). Recent studies have shown that histone lysine methylation is directly reversed by several histone demethylase families (5).Lysine methylation has also been identified in non-histone proteins that include ribulose 1,5-bisphosphate carboxylase/ oxygenase in plants (6), cytochrome c in yeast (7), mammalian TAF10 and p53 (8 -10), and ribosomal proteins in a diverse range of species (11). The methylation of ribosomal proteins has been observed in both prokaryotes and eukaryotes. In the budding yeast Saccharomyces cerevisiae, a combination of in vivo labeling and direct mass spectrometric analysis of the ribosomal proteins revealed that six of them, Rpl1, Rpl3, Rpl12, Rpl23, Rpl42, and Rpl43, are post-translationally methylated (12, 13). By analyzing the methylation state of S. cerevisiae mutant strains with deletions in candidate SET domain-containing genes, tw...

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Section: Discussionmentioning

confidence: 99%

Methylation of Ribosomal Protein L42 Regulates Ribosomal Function and Stress-adapted Cell Growth

Shirai

Sadaie

Shinmyozu³

et al. 2010

Journal of Biological Chemistry

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“…tRNA derivatives with oxidized 3′-terminal ribose were fruitfully used for studying contacts of 3′-terminus of tRNA with aminoacyl-tRNA synthetases [28][29][30][31] and human ribosomes [32,33]. In this study, we apply a set of mRNA analogs with 3′-oxidized ribose including derivatives of short oligoribonucleotides and of hepatitis C virus RNA internal ribosome entry site (HCV IRES) to reveal molecular contacts of mRNA riboses with ribosomal proteins in the human ribosome.…”

Section: Introductionmentioning

confidence: 99%

Molecular contacts of ribose-phosphate backbone of mRNA with human ribosome

Sharifulin

Grosheva

Bartuli

et al. 2015

Biochimica et Biophysica Acta (BBA) - Gene Regulatory Mechanism

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“…This protein was recently shown to contribute to the catalytic activity of the yeast ribosome at the elongation step of translation by promoting peptide bond formation (personal communication of Codjo Hountondji). This protein was found over expressed in almost all cancers [4][5][6] and could be a target for chemotherapeutic molecules. Further studies subsequently showed that some chemotherapeutic molecules can be used to target the eL42 protein as for example cycloheximide or lactimidomycine which inhibit eL42 protein through lysine (K53) of the catalytic site [11], with their carbonyl functions (C=O).…”

Section: Introductionmentioning

confidence: 99%

“…In order to better understand the functions played by the different actors, in the protein translation machinery, and identify the ribosomal components that contribute to this biological process, recent studies [6] indicates the existence of eL42 protein formerly known as L36AL (12kDa, pHi=10.59) in the large 60S subunit of the human ribosome. This protein was recently shown to contribute to the catalytic activity of the yeast ribosome at the elongation step of translation by promoting peptide bond formation (personal communication of Codjo Hountondji).…”

Section: Introductionmentioning

confidence: 99%

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Biological Evaluation of a Series of Thiosemicarbazones Targeting the Large Subunit Ribosomal Protein eL42 from Human 80S Ribosomes

Gbaguidi¹

2017

MOJBOC

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Background: Thiosemicarbazones are a family known as excellent chelators of enzyme cofactors or molecules with multiple biological activities including antitumor activity [1,2]. Recent studies by Hountondji's team has revealed that the large subunit ribosomal protein eL42 (molecular weight of 12 kDa, pHi: 10,59) plays a functional role during the elongation step of translation [3] and is over expressed in most cancers [4][5][6]. Method:The method used was the one developed by Hountondji et al. [7] which converts the OH functional groups at the positions 2' and 3' of adenine sugar of the CCA tRNA into a dialdehyde group by oxidation with NaIO 4 . Each aldehyde function can form a Schiff base with the amino group of the side chain of K53 residue of eL42, which results in a covalent bond after reduction with NaBH 4 (crosslinking). Results and Discussion:The results of different experiments (the radioactivity, and the Western Blot technique) performed in this study show that 4 molecules (citral-4-phenylthiosemicarbazone, piperitone-4-phenylthiosemicarbazone, the benzoin-4-phenylthiosemicarbazone and 4-chlorobenzophenone-4-phenylthiosemicarbazone) out of 29 thiosemicarbazones tested showed activity by inhibiting the crosslinking reaction between tRNAox and eL42. These results suggest that the inhibition produced by thiosemicarbazones consist in masking the amino group of the side chain of Lys-53 located in the catalytic site of protein eL42, preventing Schiff base formation and the crosslinking reaction. These results are in accordance with those of Schneider-Poetsch et al. [8] who found that certain compounds for example cycloheximide bind to the eL42 protein through their carbonyl function. The weaker effects of benzoin-4-phenylthiosemicarbazone and 4-chlorobenzophenone-4-phenylthiosemicarbazone as compared with citral-4-phenylthiosemicarbazone and piperitone-4-phenylthiosemicarbazone are likely to reflect the inaccessibility of the NH 2 of Lys-53 of eL42 to the thiocarbonyl groups (C=S) of these thiosemicarbazones because of steric hindrance. Finally, binding assays on Biacore 3000 confirmed interactions between active molecules and the human eL42 protein. Conclusion:The inhibitory action of thiosemicarbazones on the interaction between the protein eL42 and tRNAox prevents formation of a Schiff base between the ε-amino group of Lys-53 and an aldehyde group of tRNAox. The Lys-53 residue of eL42 which contributes to the catalysis of peptide bond formation that represents the key step in the biosynthesis of proteins is a target of choice for small molecule inhibitors. Therefore, thiosemicarbazones can be used to reduce the rate of protein synthesis in order to block the hyper-proliferation of tumor cells (Figure 1).

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The human large subunit ribosomal protein L36A-like contacts the CCA end of P-site bound tRNA

Cited by 30 publications

References 30 publications

Methylation of Ribosomal Protein L42 Regulates Ribosomal Function and Stress-adapted Cell Growth

Methylation of Ribosomal Protein L42 Regulates Ribosomal Function and Stress-adapted Cell Growth

Molecular contacts of ribose-phosphate backbone of mRNA with human ribosome

Biological Evaluation of a Series of Thiosemicarbazones Targeting the Large Subunit Ribosomal Protein eL42 from Human 80S Ribosomes

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