The solution structure of the guanine nucleotide exchange domain of human elongation factor 1β reveals a striking resemblance to that of EF-Ts from Escherichia coli

Pérez, Janice M.J.; Siegal, Gregg; Kriek, Jan; Hård, Karl; Dijk, Jan; Canters, Gerard W.; Möller, W.

doi:10.1016/s0969-2126(99)80027-6

Cited by 31 publications

(23 citation statements)

References 56 publications

(75 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…21,22 This has been observed in the crystal structure of the complex between eEF1A from Saccharomyces cerevisiae and a C-terminal catalytic fragment of eEF1BR 26 and also in the presence of GDP, GDP and Mg 2+ , and GDPNP (a nonhydrolyzable analogue of GTP). 29 Interestingly, the conformation that this loop adopts in the crystal structures of these complexes is significantly different from that reported for the equivalent loop in human eEF1BR previously determined in solution by NMR spectroscopy, 27 which implies mutual adaptation upon complex formation.…”

Section: Introductionmentioning

confidence: 81%

“…9 This rather unusual mechanism would then be similar to that of the antibiotic kirromycin, which is known to bind to prokaryotic EF1A (previously termed EF-Tu), 15 even though DB, unlike this antibiotic, does not prevent peptide bond formation. Subsequent experimental results led to the suggestion that eEF2 binding could be prevented by inhibiting eEF1A release from the ribosomal A-site, and two alternative scenarios were then envisaged: 14 (i) direct competition of the DB‚eEF1A complex with eEF2 for the same binding site on the 27) showing the role of eEF1A and eEF1B (red arrows) in translation elongation. Binding of GTP to eEF1A activates this elongation factor through a conformational change and triggers recruitment of aa-tRNA.…”

Section: Introductionmentioning

confidence: 99%

“…Although the basic structures of eEF1A and EF1A have been shown to be very similar 26 and the solution structure of the GEF domain of human eEF1BR has revealed a striking resemblance to that of EF1B (usually referred to as EF-Ts) from Escherichia coli, 27 recognition of eEF1A by eEF1BR is very different from that observed in the prokaryotic EF1A:EF1B (i.e., EF-Tu:EFTs) complex 28 and displays a highly unique feature: one loop of eEF1BR interacts with domain 2 of eEF1A in the region that is involved in the binding of the CCAaminoacyl end of tRNA. 21,22 This has been observed in the crystal structure of the complex between eEF1A from Saccharomyces cerevisiae and a C-terminal catalytic fragment of eEF1BR 26 and also in the presence of GDP, GDP and Mg 2+ , and GDPNP (a nonhydrolyzable analogue of GTP).…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Structural Basis for the Binding of Didemnins to Human Elongation Factor eEF1A and Rationale for the Potent Antitumor Activity of These Marine Natural Products

et al. 2004

View full text Add to dashboard Cite

Didemnins and tamandarins are closely related marine natural products with potent inhibitory effects on protein synthesis and cell viability. On the basis of available biochemical and structural evidence and results from molecular dynamics simulations, a model is proposed that accounts for the strong and selective binding of these compounds to human elongation factor eEF1A in the presence of GTP. We suggest that the p-methoxyphenyl ring of these cyclic depsipeptides is inserted into the same pocket in eEF1A that normally lodges either the 3′ terminal adenine of aminoacylated tRNA, as inferred from two prokaryotic EF-Tu‚GTP‚tRNA complexes, or the aromatic side chain of Phe/Tyr-163 from the nucleotide exchange factor eEF1BR, as observed in several X-ray crystal structures of a yeast eEF1A:eEF1BR complex. This pocket, which has a strong hydrophobic character, is formed by two protruding loops on the surface of eEF1A domain 2. Further stabilization of the bound depsipeptide is brought about by additional crucial interactions involving eEF1A domain 1 in such a way that the molecule fits snugly at the interface between these two domains. In the GDP-bound form of eEF1A, this binding site exists only as two separate halves, which accounts for the much greater affinity of didemnins for the GTP-bound form of this elongation factor. This binding mode is entirely different from those seen in the complexes of the homologous prokaryotic EF-Tu with kirromycin-type antibiotics or the cyclic thiazolyl peptide antibiotic GE2270A. Interestingly, the set of interactions used by didemnins to bind to eEF1A is also distinct from that used by eEF1BR or eEF1B , thus establishing a competition for binding to a common site that goes beyond simple molecular mimicry. The model presented here is consistent with both available biochemical evidence and known structure-activity relationships for these two classes of natural compounds and synthetic analogues and provides fertile ground for future research.

show abstract

Section: Introductionmentioning

confidence: 81%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Structural Basis for the Binding of Didemnins to Human Elongation Factor eEF1A and Rationale for the Potent Antitumor Activity of These Marine Natural Products

et al. 2004

View full text Add to dashboard Cite

show abstract

“…These results indicate that mutations in yEF-1␤ alter translation elongation, either directly or through the activity of yEF-1␣. According to the nuclear magnetic resonance structure of the C terminus of hEF-1␤, residues K120 and S121 lie at the end of a ␤-sheet opposite the loops predicted to play critical roles in guanine nucleotide exchange (28). Thus, the in vivo analysis of yEF-1␤ has yielded new insight into residues not predicted to play a critical role in the function of this protein.…”

Section: Fig 6 Suppression Of the Csmentioning

confidence: 99%

“…Mutational analysis directed to the first two residues of this cluster results in a large series of substitutions that confer conditional growth defects and severe sensitivity to translation inhibitors. The site of these mutations is predicted, based on the nuclear magnetic resonance structure of the C terminus of hEF-1␤ and the crystal structure of the prokaryotic homologs EF-Tu and EF-Ts, to lie at the end of a ␤-strand opposite the proposed functional loops for nucleotide exchange (18,28). While expression of the hEF-1␤ protein can replace the essential yeast protein in vivo, the hEF-1␦ cannot.…”

mentioning

confidence: 99%

Mutations in Elongation Factor 1β, a Guanine Nucleotide Exchange Factor, Enhance Translational Fidelity

Carr-Schmid

Valente

Loik

et al. 1999

Molecular and Cellular Biology

View full text Add to dashboard Cite

Translation elongation factor 1␤ (EF-1␤) is a member of the family of guanine nucleotide exchange factors, proteins whose activities are important for the regulation of G proteins critical to many cellular processes. EF-1␤ is a highly conserved protein that catalyzes the exchange of bound GDP for GTP on EF-1␣, a required step to ensure continued protein synthesis. In this work, we demonstrate that the highly conserved C-terminal region of Saccharomyces cerevisiae EF-1␤ is sufficient for normal cell growth. This region of yeast and metazoan EF-1␤ and the metazoan EF-1␤-like protein EF-1␦ is highly conserved. Human EF-1␤, but not human EF-1␦, is functional in place of yeast EF-1␤, even though both EF-1␤ and EF-1␦ have previously been shown to have guanine nucleotide exchange activity in vitro. Based on the sequence and functional homology, mutagenesis of two C-terminal residues identical in all EF-1␤ protein sequences was performed, resulting in mutants with growth defects and sensitivity to translation inhibitors. These mutants also enhance translational fidelity at nonsense codons, which correlates with a reduction in total protein synthesis. These results indicate the critical function of EF-1␤ in regulating EF-1␣ activity, cell growth, translation rates, and translational fidelity.Translation elongation requires the function of soluble protein factors. In eukaryotic organisms, the translation elongation factor 1 (EF-1) delivers aminoacyl-tRNAs (aa-tRNAs) to the A site of an elongating ribosome (6). EF-2 functions after peptide bond formation to translocate the peptidyl-tRNA to the P site (30). EF-3 is a third fungus-specific elongation factor (3). All three factors have a requirement for energy from either GTP or ATP. Only EF-1, however, requires a nucleotide exchange factor to maintain the pool of active nucleotide triphosphate-bound protein.EF-1 is composed of four subunits in metazoans (␣, ␤, ␥, and ␦) (6) but only three subunits in the yeast Saccharomyces cerevisiae (␣, ␤, and ␥) (31). The ␣ subunit is a classic G protein that binds aa-tRNAs in a GTP-dependent manner and delivers them to the A site of the elongating ribosome. After GTP hydrolysis, the resulting GDP remains bound to EF-1␣, and the protein is unable to reenter the elongation cycle until GTP is rebound. The EF-1␤ subunit functions as a guanine nucleotide exchange factor in vitro (38). While EF-1␤ is found associated with the ␥ subunit, the role of EF-1␥ remains unknown, although it modestly stimulates the activity of EF-1␤ in vitro (41). The ␦ subunit of metazoans also functions as a guanine nucleotide exchange factor in vitro but may function in vivo in the assembly of higher-order complexes of EF-1 with aa-tRNA synthetases (2). There is no biochemical or genetic evidence that yeasts contain an EF-1␦ subunit.The ␤ subunit of yeast EF-1 (yEF-1) is encoded by the single-copy essential gene TEF5 (14). The requirement for yEF-1␤ can be relieved by the presence of an extra copy of the gene encoding yEF-1␣ (20). The resulting yEF-1␤-deficient stra...

show abstract

High resolution crystal structure of bovine mitochondrial EF-tu in complex with GDP

Andersen

Thirup

Spremulli

et al. 2000

Journal of Molecular Biology

View full text Add to dashboard Cite

The solution structure of the guanine nucleotide exchange domain of human elongation factor 1β reveals a striking resemblance to that of EF-Ts from Escherichia coli

Cited by 31 publications

References 56 publications

Structural Basis for the Binding of Didemnins to Human Elongation Factor eEF1A and Rationale for the Potent Antitumor Activity of These Marine Natural Products

Structural Basis for the Binding of Didemnins to Human Elongation Factor eEF1A and Rationale for the Potent Antitumor Activity of These Marine Natural Products

Mutations in Elongation Factor 1β, a Guanine Nucleotide Exchange Factor, Enhance Translational Fidelity

High resolution crystal structure of bovine mitochondrial EF-tu in complex with GDP

Contact Info

Product

Resources

About