The primary structure of protein 44 from the large subunit of yeast ribosomes

Itoh, Takuzì; Wittmann-Liebold, B.

doi:10.1016/0014-5793(78)80447-5

Cited by 27 publications

(18 citation statements)

References 12 publications

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“…1). This mass corresponds to the mass of the gene product of Rpl42ab with the loss of the initiator methionine and the presence of two methyl groups (9,16). This wild-type 12,108 species was also observed when the analysis was repeated for ribosomal proteins from a number of SET domain mutant strains, including RKM2, SET2, SET3, SET4, SET5, SET6, and Yhl039w.…”

Section: Identification Of Two Rpl42ab Lysine Methyltransferases-mentioning

confidence: 94%

“…The resulting large subunit proteins were fractionated by reversephase HPLC and their intact masses determined by electrospray mass spectrometry as described under "Experimental Procedures." We focused on the large ribosomal protein Rpl42ab since evidence has been presented for monomethyllysine formation at two sites (16). The presence of two methyl groups on Rpl42ab was supported by the mass spectrometric determination of the intact protein.…”

Section: Identification Of Two Rpl42ab Lysine Methyltransferases-mentioning

confidence: 99%

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Identification of Two SET Domain Proteins Required for Methylation of Lysine Residues in Yeast Ribosomal Protein Rpl42ab

Webb

Laganowsky

Whitelegge

et al. 2008

Journal of Biological Chemistry

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We show that the Saccharomyces cerevisiae ribosomal protein Rpl42ab (the identical product of the RPL42A and RPL42B genes) is monomethylated at Lys-40 and Lys-55. The methylation of Lys-40 is dependent upon the Ybr030w gene product; the methylation of Lys-55 is dependent upon the Set7 gene product. Ybr030w and SET7 genes both encode SET domain containing proteins homologous to known protein lysine methyltransferases, suggesting that their products are the specific enzymes responsible for the monomethylation of the two sites in Rpl42ab. We thus designate Ybr030w as Rkm3 and Set7 as Rkm4. Yeast strains with deletions in both the Ybr030w and SET7 genes produce unmethylated Rpl42ab. A slow growth phenotype was seen for the SET7 deletion strain and the double knock-out when grown in low concentrations of the eukaryotic protein synthesis inhibitor, cycloheximide. These results suggest that modification of Rpl42ab at Lys-55 can fine-tune its structure to avoid inhibition. An intact mass fragmentation approach ("top down mass spectrometry") was used to quantitate the extent of methylation of Rpl42ab. In wild-type strains, it was found that 78% was monomethylated at both Lys-40 and Lys-55 and that 22% was a mixture of species with either Lys-40 or Lys-55 monomethylated. The top down approach was also used to reevaluate the methylation sites of Rpl12ab. We found that the yeast Rpl12ab protein is dimethylated at the N-terminal proline residue, trimethylated at Lys-3 by Rkm2, and monomethylated at Arg-66.The emergence of histone protein lysine methyltransferases that regulate the epigenetic state of chromatin highlights the biological importance of this modification (1, 2). Protein lysine methylation has also been found to occur in over a hundred proteins ranging from flagellin in Salmonella typhimurium to RUBISCO in plants to transcription initiation factor TAFT10 in Homo sapiens (3). Physiological functions of lysine methylation of non-histone proteins remain unclear, and the enzymes responsible for the modifications have only recently begun to be characterized.With the exceptions of the yeast DOT1 and the bacterial PrmA enzymes, known protein lysine methyltransferases all belong to the family of SET domain-containing proteins (4 -6). Structurally the SET domain enzymes are distinct from the more common seven ␤-strand class of methyltransferases. All SET domain methyltransferases studied to date appear to only catalyze protein lysine methylation reactions (3-6). These enzymes are capable of adding up to three methyl groups to the side chain amino group of lysine residues to form mono-, di-, and trimethylated derivatives. Interestingly, this modification can be reversed by members of a large family of demethylases (7).We have been interested in identifying protein lysine methyltransferases that are involved in the formation of methylation marks on ribosomal proteins. Multiple proteins in both the small and large ribosomal subunits have been shown to be modified at lysine residues in both prokaryotes and eukaryotes (8). Our...

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Section: Identification Of Two Rpl42ab Lysine Methyltransferases-mentioning

confidence: 94%

Section: Identification Of Two Rpl42ab Lysine Methyltransferases-mentioning

confidence: 99%

Identification of Two SET Domain Proteins Required for Methylation of Lysine Residues in Yeast Ribosomal Protein Rpl42ab

Webb

Laganowsky

Whitelegge

et al. 2008

Journal of Biological Chemistry

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“…2a identical to each other, but for some unknown reason they Isolation and phenotype of L41-related protein genes from a showed a few amino acid residues that were different from few other yeasts. To identify the specific amino acid resithe published sequence (5). The homology of the deduced due(s) responsible for the difference in CYH sensitivity amino acid sequences of the CYH-sensitive L41 protein and between these two L41-related ribosomal proteins, a clone the CYH-resistant RIM-C protein was about 85% (Fig.…”

Section: Methodsmentioning

confidence: 99%

Drastic alteration of cycloheximide sensitivity by substitution of one amino acid in the L41 ribosomal protein of yeasts

et al. 1992

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Cycloheximide is one of the antibiotics that inhibit protein synthesis in most eukaryotic cells. We have found that a yeast, Candida maltosa, is resistant to the drug because it possesses a cycloheximide-resistant ribosome, and we have isolated the gene responsible for this. In this study, we sequenced this gene and found that the gene encodes a protein homologous to the L41 ribosomal protein of Saccharomyces cerevisiae, whose amino acid sequence has already been reported. Two genes for L41 protein, named L41a and L41b, independently present in the genome of S. cerevisiae, were isolated. L41-related genes were also isolated from a few other yeast species. Each of these genes has an intron at the same site of the open reading frame. Comparison of their deduced amino acid sequences and their ability to confer cycloheximide resistance to S. cerevisiae, when introduced in a high-copy-number plasmid, suggested that the 56th amino acid residue of the L41 protein determines the sensitivity of the ribosome to cycloheximide; the amino acid is glutamine in the resistant ribosome, whereas that in the sensitive ribosome is proline. This was confirmed by constructing a cycloheximide-resistant strain of S. cerevisiae having a disrupted L41a gene and an L41b gene with a substitution of the glutamine codon for the proline codon.

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“…ORF A was highly similar (75% identity) to four known eukaryotic ribosomal proteins: S. cerevisiue rp44 (also called L41) (Itoh and Wittmann-Liebold, 1978), human L44 (Davies et al, 1986), rat L36a (Gallagher et d., 1988) and Trypanosoma brucei L 44 (Tebabi et al, 1990). In addition, two other DNA sequences, not available in data banks, encoded proteins similar to ORF A: Schwunniomyces occidentulis (also known as Sw. custellii;del Pozo, et al, 1993) and Cundidu multosu (Takagi et al, 1989).…”

Section: Orf a Encodes Ribosomal Protein L41mentioning

confidence: 97%

Natural cycloheximide resistance in yeast

Dehoux¹,

Davies²,

Cannon

1993

European Journal of Biochemistry

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The yest Kluyveromyces lactis is resistant to high concentrations (1 mg/ml) of the antibiotic cycloheximide. Using in vitro translation studies it was confirmed that this extreme resistance is a property of ribosomes. The resistance determinant from K. lactis was cloned into Saccharomyces cerevisiae. Nucleotide sequence analysis of the determinant demonstrated that resistance was conferred by the K. Lactis ribosomal protein L41. K. lactis was shown to contain only one copy of the gene that encodes this protein and the gene was located to chromosome III. In contrast, S. cerevisiae was found to contain multiple copies of the gene for the corresponding ribosomal protein L41 which mapped to two of the three chromosomes V, XIV and VIII. Since the cycloheximide‐resistance gene of K. lactis causes essentially complete protection against inhibition by the drug, it is likely to be particularly useful as a selective marker in eukaryotic gene transfer studies.

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The primary structure of protein 44 from the large subunit of yeast ribosomes

Cited by 27 publications

References 12 publications

Identification of Two SET Domain Proteins Required for Methylation of Lysine Residues in Yeast Ribosomal Protein Rpl42ab

Identification of Two SET Domain Proteins Required for Methylation of Lysine Residues in Yeast Ribosomal Protein Rpl42ab

Drastic alteration of cycloheximide sensitivity by substitution of one amino acid in the L41 ribosomal protein of yeasts

Natural cycloheximide resistance in yeast

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