The nucleotide sequence of the gene coding for the elongation factor 1α in Sulfolobus solfataricus. Homology of the product with related proteins

Arcari, Paolo; Gallo, Monica; Ianniciello, G.; Russo, Antonio Dello; Bocchini, Vincenzo

doi:10.1016/0167-4781(94)90296-8

Cited by 15 publications

(14 citation statements)

References 39 publications

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“…Moreover, we used the 3D structure of EF-1A GDP from Sulfolobus solfataricus as a template for the prediction of phosphorylation putative sites. 26 To conclude, in the human epidermoid lung cancer cells, H1355, the upregulation of eEF-1A intracellular content mediated by IFNa is associated with an increase in the phosphorylation of serine and threonine residues of the protein. Our findings indicate that the effect is mediated by C-Raf, and appears to play a pro-survival function.…”

Section: Discussionmentioning

confidence: 83%

C-Raf antagonizes apoptosis induced by IFN-α in human lung cancer cells by phosphorylation and increase of the intracellular content of elongation factor 1A

Lamberti

Longo

Marra³

et al. 2007

Cell Death Differ

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Interferon a (IFNa) induces both apoptosis and a counteracting epidermal growth factor Erk-dependent survival response in cancer cells. In this report, IFNa increased eukaryotic elongation factor 1A (eEF-1A) protein expression by inhibition of eEF-1A degradation via a proteasome-dependent pathway. The reduction of the expression level of eEF-1A by RNA interference enhanced the apoptosis induced by IFNa on the same cells. Moreover, IFNa induced the phosphorylation of both serine and threonine in eEF-1A. These effects were paralleled by an increased co-immunoprecipitation and colocalization of eEF-1A with C-Raf. The suppression of C-Raf kinase activity with the inhibitor BAY 43-9006 completely antagonized the increase of both eEF-1A phosphorylation and expression and of C-Raf/eEF-1A colocalization induced by IFNa and enhanced apoptosis and eEF-1A ubiquitination. Cell transfection with the mutated K48R ubiquitin increased EF-1A expression and desensitized tumor cells to the modulating effects of IFNa. The dynamic simulation of 3Dstructure of eEF-1A identified putative serine and threonine phosphorylation sites. In conclusion, the interaction between eEF-1A and C-Raf increases eEF-1A stability and induces a survival activity.

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Section: Discussionmentioning

confidence: 83%

C-Raf antagonizes apoptosis induced by IFN-α in human lung cancer cells by phosphorylation and increase of the intracellular content of elongation factor 1A

Lamberti

Longo

Marra³

et al. 2007

Cell Death Differ

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“…Moreover, the GTPase activity of eEF1A is also used to enhance the accuracy of codon recognition. 4, 5 The functions of eEF1A in the elongation cycle have been extensively investigated in eubacteria, for example, Escherichia coli (EF-Tu) as well as in archaea, for example, Sulfolobus sulfataricus 6, 7 and eukaryotes, for example, Saccharomyces cerevisiae . These studies showed the high conservation of the elongation factor between the three kingdoms as well as among a wide range of different species confirming its importance for life.…”

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confidence: 99%

Raf kinases mediate the phosphorylation of eukaryotic translation elongation factor 1A and regulate its stability in eukaryotic cells

et al. 2012

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We identified eukaryotic translation elongation factor 1A (eEF1A) Raf-mediated phosphorylation sites and defined their role in the regulation of eEF1A half-life and of apoptosis of human cancer cells. Mass spectrometry identified in vitro S21 and T88 as phosphorylation sites mediated by B-Raf but not C-Raf on eEF1A1 whereas S21 was phosphorylated on eEF1A2 by both B- and C-Raf. Interestingly, S21 belongs to the first eEF1A GTP/GDP-binding consensus sequence. Phosphorylation of S21 was strongly enhanced when both eEF1A isoforms were preincubated prior the assay with C-Raf, suggesting that the eEF1A isoforms can heterodimerize thus increasing the accessibility of S21 to the phosphate. Overexpression of eEF1A1 in COS 7 cells confirmed the phosphorylation of T88 also in vivo. Compared with wt, in COS 7 cells overexpressed phosphodeficient (A) and phospho-mimicking (D) mutants of eEF1A1 (S21A/D and T88A/D) and of eEF1A2 (S21A/D), resulted less stable and more rapidly proteasome degraded. Transfection of S21 A/D eEF1A mutants in H1355 cells increased apoptosis in comparison with the wt isoforms. It indicates that the blockage of S21 interferes with or even supports C-Raf induced apoptosis rather than cell survival. Raf-mediated regulation of this site could be a crucial mechanism involved in the functional switching of eEF1A between its role in protein biosynthesis and its participation in other cellular processes.

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“…The alignment of the nucleotide sequence of the Ss MT3EF‐1α gene (EMBL accession number AJ312397) with the corresponding sequence of the gene isolated from MT4 strain[6] showed the presence of three base substitutions, namely G 43 →A, G 720 →C and C 954 →T as numbered from the start codon (not shown). The G 43 →A change led to a different amino acid residue (Ile15 instead of Val15 in Ss MT4EF‐1α) whereas the other base differences led to synonymous codons.…”

Section: Resultsmentioning

confidence: 99%

“…We have previously reported the purification and characterisation of the translational elongation factor 1α (EF‐1α) from the archaeon S. solfataricus , strain MT4 ( Ss MT4EF‐1α) (ATCC 49255)[5] together with the cloning and the structural organisation of its encoding gene [6,7]. Because EF‐1α is present in all the living organisms, it was of interest to analyse the molecular properties of the enzyme from the MT3 strain ( Ss MT3EF‐1α) and to compare them with those of the MT4 enzyme.…”

Section: Introductionmentioning

confidence: 99%

Cloning, expression and evolution of the gene encoding the elongation factor 1Î± from a low thermophilicSulfolobus solfataricusstrain

Cantiello

Lamberti

Longo

et al. 2003

FEMS Microbiology Letters

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The gene encoding the elongation factor 1alpha (EF-1alpha) from the archaeon Sulfolobus solfataricus strain MT3 (optimum growth temperature 75 degrees C) was cloned, sequenced and expressed in Escherichia coli. The structural and biochemical properties of the purified enzyme were compared to those of EF-1alpha isolated from S. solfataricus strain MT4 (optimum growth temperature 87 degrees C). Only one amino acid change (Val15-->Ile) was found. Interestingly, the difference was in the first guanine nucleotide binding consensus sequence G(13)HIDHGK and was responsible for a reduced efficiency in protein synthesis, which was accompanied by an increased affinity for both guanosine diphosphate (GDP) and guanosine triphosphate (GTP), and an increased efficiency in the intrinsic GTPase activity. Despite the different thermophilicities of the two microorganisms, only very marginal effects on the thermal properties of the enzyme were observed. Molecular evolution among EF-1alpha genes from Sulfolobus species showed that the average rate of nucleotide substitution per site per year (0.0312x10(-9)) is lower than that reported for other functional genes.

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The nucleotide sequence of the gene coding for the elongation factor 1α in Sulfolobus solfataricus. Homology of the product with related proteins

Cited by 15 publications

References 39 publications

C-Raf antagonizes apoptosis induced by IFN-α in human lung cancer cells by phosphorylation and increase of the intracellular content of elongation factor 1A

C-Raf antagonizes apoptosis induced by IFN-α in human lung cancer cells by phosphorylation and increase of the intracellular content of elongation factor 1A

Raf kinases mediate the phosphorylation of eukaryotic translation elongation factor 1A and regulate its stability in eukaryotic cells

Cloning, expression and evolution of the gene encoding the elongation factor 1Î± from a low thermophilicSulfolobus solfataricusstrain

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