Differential transcription of the two Saccharomyces cerevisiae genes encoding elongation factor 2

Gene ◽  
1994 ◽  
Vol 148 (1) ◽  
pp. 143-147 ◽  
Author(s):  
Sarah Veldman ◽  
Sulekha Rao ◽  
James W. Bodley
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Elongation Factor ◽  
Elongation Factor 2 ◽  
Genes Encoding ◽  
Differential Transcription

scholarly journals Saccharomyces cerevisiae elongation factor 2. Genetic cloning, characterization of expression, and G-domain modeling.

1992 ◽  
Vol 267 (2) ◽  
pp. 1190-1197 ◽  
Author(s):  
J P Perentesis ◽  
L D Phan ◽  
W B Gleason ◽  
D C LaPorte ◽  
D M Livingston ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Elongation Factor ◽  
Domain Modeling ◽  
Elongation Factor 2

Elongation factor EF-1 alpha gene dosage alters translational fidelity in Saccharomyces cerevisiae

1989 ◽  
Vol 9 (10) ◽  
pp. 4571-4575
Author(s):  
J M Song ◽  
S Picologlou ◽  
C M Grant ◽  
M Firoozan ◽  
M F Tuite ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Gene Dosage ◽  
Elongation Factor ◽  
Nonsense Suppression ◽  
Translational Fidelity ◽  
Genes Encoding ◽  
Alpha Gene ◽  
Nonsense Codons

Changes in the dosage of genes encoding elongation factor EF-1 alpha were shown to cause parallel changes in the misreading of nonsense codons. Higher amounts of EF-1 alpha were correlated with increased nonsense suppression, suggesting that the level of EF-1 alpha is critically involved in translational fidelity.

scholarly journals Diphtheria toxin-resistant mutants of Saccharomyces cerevisiae.

1985 ◽  
Vol 5 (12) ◽  
pp. 3357-3360 ◽  
Author(s):  
J Y Chen ◽  
J W Bodley ◽  
D M Livingston
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Diphtheria Toxin ◽  
Elongation Factor ◽  
Selection Procedure ◽  
Prolonged Storage ◽  
Elongation Factor 2 ◽  
Resistant Phenotype ◽  
Complementation Groups ◽  
Mendelian Character

We developed a selection procedure based on the observation that diphtheria toxin kills spheroplasts of Saccharomyces cerevisiae (Murakami et al., Mol. Cell. Biol. 2:588-592, 1982); this procedure yielded mutants resistant to the in vitro action of the toxin. Spheroplasts of mutagenized S. cerevisiae were transformed in the presence of diphtheria toxin, and the transformed survivors were screened in vitro for toxin-resistant elongation factor 2. Thirty-one haploid ADP ribosylation-negative mutants comprising five complementation groups were obtained by this procedure. The mutants grew normally and were stable to prolonged storage. Heterozygous diploids produced by mating wild-type sensitive cells with the mutants revealed that in each case the resistant phenotype was recessive to the sensitive phenotype. Sporulation of these diploids yielded tetrads in which the resistant phenotype segregated as a single Mendelian character. From these observations, we concluded that these mutants are defective in the enzymatic steps responsible for the posttranslational modification of elongation factor 2 which is necessary for recognition by diphtheria toxin.

scholarly journals Elongation factor EF-1 alpha gene dosage alters translational fidelity in Saccharomyces cerevisiae.

1989 ◽  
Vol 9 (10) ◽  
pp. 4571-4575 ◽  
Author(s):  
J M Song ◽  
S Picologlou ◽  
C M Grant ◽  
M Firoozan ◽  
M F Tuite ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Gene Dosage ◽  
Elongation Factor ◽  
Nonsense Suppression ◽  
Translational Fidelity ◽  
Genes Encoding ◽  
Alpha Gene ◽  
Nonsense Codons

Changes in the dosage of genes encoding elongation factor EF-1 alpha were shown to cause parallel changes in the misreading of nonsense codons. Higher amounts of EF-1 alpha were correlated with increased nonsense suppression, suggesting that the level of EF-1 alpha is critically involved in translational fidelity.

Diphtheria toxin-resistant mutants of Saccharomyces cerevisiae

1985 ◽  
Vol 5 (12) ◽  
pp. 3357-3360
Author(s):  
J Y Chen ◽  
J W Bodley ◽  
D M Livingston
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Diphtheria Toxin ◽  
Elongation Factor ◽  
Selection Procedure ◽  
Prolonged Storage ◽  
Elongation Factor 2 ◽  
Resistant Phenotype ◽  
Complementation Groups ◽  
Mendelian Character

We developed a selection procedure based on the observation that diphtheria toxin kills spheroplasts of Saccharomyces cerevisiae (Murakami et al., Mol. Cell. Biol. 2:588-592, 1982); this procedure yielded mutants resistant to the in vitro action of the toxin. Spheroplasts of mutagenized S. cerevisiae were transformed in the presence of diphtheria toxin, and the transformed survivors were screened in vitro for toxin-resistant elongation factor 2. Thirty-one haploid ADP ribosylation-negative mutants comprising five complementation groups were obtained by this procedure. The mutants grew normally and were stable to prolonged storage. Heterozygous diploids produced by mating wild-type sensitive cells with the mutants revealed that in each case the resistant phenotype was recessive to the sensitive phenotype. Sporulation of these diploids yielded tetrads in which the resistant phenotype segregated as a single Mendelian character. From these observations, we concluded that these mutants are defective in the enzymatic steps responsible for the posttranslational modification of elongation factor 2 which is necessary for recognition by diphtheria toxin.

scholarly journals A Chemical Genomic Screen in Saccharomyces cerevisiae Reveals a Role for Diphthamidation of Translation Elongation Factor 2 in Inhibition of Protein Synthesis by Sordarin

2008 ◽  
Vol 52 (5) ◽  
pp. 1623-1629 ◽  
Author(s):  
Javier Botet ◽  
María Rodríguez-Mateos ◽  
Juan P. G. Ballesta ◽  
José Luis Revuelta ◽  
Miguel Remacha
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Protein Synthesis ◽  
Elongation Factor ◽  
Translation Elongation Factor ◽  
Cellular Mechanisms ◽  
Translation Elongation ◽  
Chemical Genomic ◽  
Elongation Factor 2 ◽  
Fungal Organisms ◽  
Inhibitor Of Protein Synthesis

ABSTRACT Sordarin and its derivatives are antifungal compounds of potential clinical interest. Despite the highly conserved nature of the fungal and mammalian protein synthesis machineries, sordarin is a selective inhibitor of protein synthesis in fungal organisms. In cells sensitive to sordarin, its mode of action is through preventing the release of translation elongation factor 2 (eEF2) during the translocation step, thus blocking protein synthesis. To further investigate the cellular components required for the effects of sordarin in fungal cells, we have used the haploid deletion collection of Saccharomyces cerevisiae to systematically identify genes whose deletion confers sensitivity or resistance to the compound. Our results indicate that genes in a number of cellular pathways previously unknown to play a role in sordarin response are involved in its growth effects on fungal cells and reveal a specific requirement for the diphthamidation pathway of cells in causing eEF2 to be sensitive to the effects of sordarin on protein synthesis. Our results underscore the importance of the powerful genomic tools developed in yeast (Saccharomyces cerevisiae) to more comprehensively understanding the cellular mechanisms involved in the response to therapeutic agents.

scholarly journals DPH5, a methyltransferase gene required for diphthamide biosynthesis in Saccharomyces cerevisiae.

1992 ◽  
Vol 12 (9) ◽  
pp. 4026-4037 ◽  
Author(s):  
L C Mattheakis ◽  
W H Shen ◽  
R J Collier
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Sequence Similarity ◽  
Elongation Factor ◽  
Complementation Group ◽  
Sequence Motifs ◽  
Null Mutant ◽  
New Family ◽  
Elongation Factor 2 ◽  
Intracellular Expression ◽  
High Concentrations

A mutant of Saccharomyces cerevisiae defective in the S-adenosylmethionine (AdoMet)-dependent methyltransferase step of diphthamide biosynthesis was selected by intracellular expression of the F2 fragment of diphtheria toxin (DT) and shown to belong to complementation group DPH5. The DPH5 gene was cloned, sequenced, and found to encode a 300-residue protein with sequence similarity to bacterial AdoMet:uroporphyrinogen III methyltransferases, enzymes involved in cobalamin (vitamin B12) biosynthesis. Both DPH5 and AdoMet:uroporphyrinogen III methyltransferases lack sequence motifs commonly found in other methyltransferases and may represent a new family of AdoMet:methyltransferases. The DPH5 protein was produced in Escherichia coli and shown to be active in methylation of elongation factor 2 partially purified from the dph5 mutant. A null mutation of the chromosomal DPH5 gene did not affect cell viability, in agreement with other studies indicating that diphthamide is not required for cell survival. The dph5 null mutant survived expression of three enzymically attenuated DT fragments but was killed by expression of fully active DT fragment A. Consistent with these results, elongation factor 2 from the dph5 null mutant was found to have weak ADP-ribosyl acceptor activity, which was detectable only in the presence of high concentrations of fragment A.

scholarly journals Translational Roles of Elongation Factor 2 Protein Lysine Methylation

2014 ◽  
Vol 289 (44) ◽  
pp. 30511-30524 ◽  
Author(s):  
Maria C. Dzialo ◽  
Kyle J. Travaglini ◽  
Sean Shen ◽  
Kevin Roy ◽  
Guillaume F. Chanfreau ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Protein Synthesis ◽  
Elongation Factor ◽  
Amino Acid Sequences ◽  
Fine Tuning ◽  
Lysine Methylation ◽  
Translational Machinery ◽  
Elongation Factor 2 ◽  
Translational Apparatus

Methylation of various components of the translational machinery has been shown to globally affect protein synthesis. Little is currently known about the role of lysine methylation on elongation factors. Here we show that in Saccharomyces cerevisiae, the product of the EFM3/YJR129C gene is responsible for the trimethylation of lysine 509 on elongation factor 2. Deletion of EFM3 or of the previously described EFM2 increases sensitivity to antibiotics that target translation and decreases translational fidelity. Furthermore, the amino acid sequences of Efm3 and Efm2, as well as their respective methylation sites on EF2, are conserved in other eukaryotes. These results suggest the importance of lysine methylation modification of EF2 in fine tuning the translational apparatus.

DPH5, a methyltransferase gene required for diphthamide biosynthesis in Saccharomyces cerevisiae

1992 ◽  
Vol 12 (9) ◽  
pp. 4026-4037
Author(s):  
L C Mattheakis ◽  
W H Shen ◽  
R J Collier
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Sequence Similarity ◽  
Elongation Factor ◽  
Complementation Group ◽  
Sequence Motifs ◽  
Null Mutant ◽  
New Family ◽  
Elongation Factor 2 ◽  
Intracellular Expression ◽  
High Concentrations

A mutant of Saccharomyces cerevisiae defective in the S-adenosylmethionine (AdoMet)-dependent methyltransferase step of diphthamide biosynthesis was selected by intracellular expression of the F2 fragment of diphtheria toxin (DT) and shown to belong to complementation group DPH5. The DPH5 gene was cloned, sequenced, and found to encode a 300-residue protein with sequence similarity to bacterial AdoMet:uroporphyrinogen III methyltransferases, enzymes involved in cobalamin (vitamin B12) biosynthesis. Both DPH5 and AdoMet:uroporphyrinogen III methyltransferases lack sequence motifs commonly found in other methyltransferases and may represent a new family of AdoMet:methyltransferases. The DPH5 protein was produced in Escherichia coli and shown to be active in methylation of elongation factor 2 partially purified from the dph5 mutant. A null mutation of the chromosomal DPH5 gene did not affect cell viability, in agreement with other studies indicating that diphthamide is not required for cell survival. The dph5 null mutant survived expression of three enzymically attenuated DT fragments but was killed by expression of fully active DT fragment A. Consistent with these results, elongation factor 2 from the dph5 null mutant was found to have weak ADP-ribosyl acceptor activity, which was detectable only in the presence of high concentrations of fragment A.

scholarly journals Functional and physical interactions of Krr1p, a Saccharomyces cerevisiae nucleolar protein.

2004 ◽  
Vol 51 (1) ◽  
pp. 173-187 ◽  
Author(s):  
Robert Gromadka ◽  
Iwona Karkusiewicz ◽  
Bozenna Rempoła ◽  
Joanna Rytka
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Ribosomal Proteins ◽  
Northern Blot Analysis ◽  
Affinity Purification ◽  
Elongation Factor ◽  
Translation Elongation ◽  
Multicopy Suppressors ◽  
Genes Encoding ◽  
Cold Sensitive ◽  
Physical Interactions

The Krr1 protein of Saccharomyces cerevisiae is involved in processing of pre-rRNA and assembly of pre-ribosomal 40S subunits. To further investigate the function of Krr1p we constructed a conditional cold sensitive mutant krr1-21, and isolated seven genes from Schizosaccharomyces pombe whose products suppressed the cold sensitive phenotype of krr1-21 cells. Among the multicopy suppressors we found genes coding for translation elongation factor EF-1alpha, a putative ribose methyltransferase and five genes encoding ribosomal proteins. Using the tandem affinity purification (TAP) method we identified thirteen S. cerevisiae ribosomal proteins interacting with Krr1p. Taken together, these results indicate that Krr1p interacts functionally as well as physically with ribosomal proteins. Northern blot analysis revealed that changes in the level of krr1-21 mRNA were accompanied by similar changes in the level of mRNAs of genes encoding ribosomal proteins. Thus, Krr1p and the genes encoding ribosomal proteins it interacts with seem to be coordinately regulated at the level of transcription.

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