Methylated proteins and amino acids in the ribosomes of Saccharomyces cerevisiae

Lhoest, Jacques; Lobet, Yves; Costers, Eric; Colson, Charles

doi:10.1111/j.1432-1033.1984.tb08233.x

Cited by 33 publications

(35 citation statements)

References 23 publications

(18 reference statements)

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“…In fact, asymmetric dimethylarginine is the predominant methylated amino acid in both the eukaryotic 40S and 60S ribosomal subunits (15). Arginine methylation of ribosomal proteins is evolutionarily conserved among eukaryotes (44,51,78) and fluctuates during the cell cycle (14). Ribosomal proteins can also be modified by lysine methylation, and methyltransferases that catalyze such modifications in ribosomal proteins were recently identified in eukaryotes (75,76).…”

Section: Protein Arginine Methyltransferasementioning

confidence: 99%

Protein Arginine Methyltransferases: from Unicellular Eukaryotes to Humans

2007

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Section: Protein Arginine Methyltransferasementioning

confidence: 99%

Protein Arginine Methyltransferases: from Unicellular Eukaryotes to Humans

2007

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“…Methylation and phosphorylation are the two most common posttranslational protein modifications observed on ribosomal proteins. Methylation has been previously observed on lysine and͞or arginine residues in a number of yeast large ribosomal proteins (9,21,22); however, the low reported stoichiometry of methylation (22), and differential modification by multiple methylation enzymes (47), may have led to different subsets of methylated proteins having been identified previously. Consistent with Lhoest and coworkers (21), we observe sample components consistent with mono-methylation of rpL3 and multiple methylations of rpL12AB and rpL23AB.…”

Section: N-terminal Modificationsmentioning

confidence: 99%

“…There are currently 137 established ribosomal protein genes in Saccharomyces cerevisiae, which encode 79 unique ribosomal proteins (46 large subunit and 33 small subunit) (8). The physical and biochemical characterization of yeast ribosomes has been complicated not only by the large number of proteins involved, and their highly basic amino acid content, but also by the presence of modifications, often in combination, including N-terminal processing and acetylation (9), methylation (21,22), and phosphorylation (23,24).…”

mentioning

confidence: 99%

Direct mass spectrometric analysis of intact proteins of the yeast large ribosomal subunit using capillary LC/FTICR

Lee

Berger

Martinović

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

182

133

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Electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry coupled with capillary reverse-phase liquid chromatography was used to characterize intact proteins from the large subunit of the yeast ribosome. High mass measurement accuracy, achieved by ''mass locking'' with an internal standard from a dual electrospray ionization source, allowed identification of ribosomal proteins. Analyses of the intact proteins revealed information on cotranslational and posttranslational modifications of the ribosomal proteins that included loss of the initiating methionine, acetylation, methylation, and proteolytic maturation. High-resolution separations permitted differentiation of protein isoforms having high structural similarity as well as proteins from their modified forms, facilitating unequivocal assignments. The study identified 42 of the 43 core large ribosomal subunit proteins and 58 (of 64 possible) core large subunit protein isoforms having unique masses in a single analysis. These results demonstrate the basis for the high-throughput analyses of complex mixtures of intact proteins, which we believe will be an important complement to other approaches for defining protein modifications and their changes resulting from physiological processes or environmental perturbations.M ass spectrometry has evolved into a powerful tool for analyzing biomolecules because of the development of matrix-assisted laser desorption ionization (1, 2) and electrospray ionization (ESI) (3) and advances in both mass analyzers and data processing capabilities. With these ionization methods, which are amenable to megadalton-size molecules, the broadly useful detection and characterization of biopolymers by mass spectrometric methods are feasible. For example, MS has become a preferred analytical tool for proteome analyses, in which dynamic populations of proteins are identified and quantified. Proteins are now routinely identified by mass spectrometric and tandem mass spectrometric analysis of proteolytic digests of individual protein spots on two-dimensional (2D) PAGE (4). However, the intact protein level analyses of complex protein mixtures, while potentially providing complementary and direct information for protein identification, have been a much greater experimental challenge and only rarely attempted (5, 6).Many pivotal cellular processes are not carried out by individual proteins, but rather by large protein-protein complexes and protein-nucleic acid complexes. The analyses of such complexes have been greatly expanded by improvements in both biological separations and MS. One of the more complicated protein complexes yet studied by MS is the cytosolic ribosome complex. Efforts to define the protein composition (7, 8), modifications (9), and subunit interactions (10) of ribosomes generally have mirrored the development of biomolecular analysis techniques and experimental approaches for the study of noncovalent protein complexes.Ribosomes are the canonical example of ribonucleoprotein complexes in both prokaryot...

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“…In Escherichia coli, methylation of ribosomal protein L3 by the PrmB methyltransferase has been shown to modulate ribosome biogenesis (Lhoest and Colson 1981;Lhoest et al 1984). Escherichia coli L11 contains nine methyl groups in close proximity to the GTPase-associated center (GAC) and the binding site of translation factors, suggesting that they could play a role in translation (Dognin and WittmannLiebold 1980).…”

Section: Introductionmentioning

confidence: 99%

Methylation of yeast ribosomal protein Rpl3 promotes translational elongation fidelity

Al-Hadid

Roy

Chanfreau

et al. 2016

RNA

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Rpl3, a highly conserved ribosomal protein, is methylated at histidine 243 by the Hpm1 methyltransferase in Saccharomyces cerevisiae. Histidine 243 lies close to the peptidyl transferase center in a functionally important region of Rpl3 designated as the basic thumb that coordinates the decoding, peptidyl transfer, and translocation steps of translation elongation. Hpm1 was recently implicated in ribosome biogenesis and translation. However, the biological role of methylation of its Rpl3 substrate has not been identified. Here we interrogate the role of Rpl3 methylation at H243 by investigating the functional impact of mutating this histidine residue to alanine (rpl3-H243A). Akin to Hpm1-deficient cells, rpl3-H243A cells accumulate 35S and 23S pre-rRNA precursors to a similar extent, confirming an important role for histidine methylation in pre-rRNA processing. In contrast, Hpm1-deficient cells but not rpl3-H243A mutants show perturbed levels of ribosomal subunits. We show that Hpm1 has multiple substrates in different subcellular fractions, suggesting that methylation of proteins other than Rpl3 may be important for controlling ribosomal subunit levels. Finally, translational fidelity assays demonstrate that like Hpm1-deficient cells, rpl3-H243A mutants have defects in translation elongation resulting in decreased translational accuracy. These data suggest that Rpl3 methylation at H243 is playing a significant role in translation elongation, likely via the basic thumb, but has little impact on ribosomal subunit levels. Hpm1 is therefore a multifunctional methyltransferase with independent roles in ribosome biogenesis and translation.

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Methylated proteins and amino acids in the ribosomes of Saccharomyces cerevisiae

Cited by 33 publications

References 23 publications

Protein Arginine Methyltransferases: from Unicellular Eukaryotes to Humans

Protein Arginine Methyltransferases: from Unicellular Eukaryotes to Humans

Direct mass spectrometric analysis of intact proteins of the yeast large ribosomal subunit using capillary LC/FTICR

Methylation of yeast ribosomal protein Rpl3 promotes translational elongation fidelity

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