Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Hypusine-containing protein eIF5A promotes translation elongation


Translation elongation factors facilitate protein synthesis by the ribosome. Previous studies identified two universally conserved translation elongation factors, EF-Tu in bacteria (known as eEF1A in eukaryotes) and EF-G (eEF2), which deliver aminoacyl-tRNAs to the ribosome and promote ribosomal translocation, respectively1. The factor eIF5A (encoded by HYP2 and ANB1 in Saccharomyces cerevisiae), the sole protein in eukaryotes and archaea to contain the unusual amino acid hypusine (Nε-(4-amino-2-hydroxybutyl)lysine)2, was originally identified based on its ability to stimulate the yield (endpoint) of methionyl-puromycin synthesis—a model assay for first peptide bond synthesis thought to report on certain aspects of translation initiation3,4. Hypusine is required for eIF5A to associate with ribosomes5,6 and to stimulate methionyl-puromycin synthesis7. Because eIF5A did not stimulate earlier steps of translation initiation8, and depletion of eIF5A in yeast only modestly impaired protein synthesis9, it was proposed that eIF5A function was limited to stimulating synthesis of the first peptide bond or that eIF5A functioned on only a subset of cellular messenger RNAs. However, the precise cellular role of eIF5A is unknown, and the protein has also been linked to mRNA decay, including the nonsense-mediated mRNA decay pathway10,11, and to nucleocytoplasmic transport12,13. Here we use molecular genetic and biochemical studies to show that eIF5A promotes translation elongation. Depletion or inactivation of eIF5A in the yeast S. cerevisiae resulted in the accumulation of polysomes and an increase in ribosomal transit times. Addition of recombinant eIF5A from yeast, but not a derivative lacking hypusine, enhanced the rate of tripeptide synthesis in vitro. Moreover, inactivation of eIF5A mimicked the effects of the eEF2 inhibitor sordarin, indicating that eIF5A might function together with eEF2 to promote ribosomal translocation. Because eIF5A is a structural homologue of the bacterial protein EF-P14,15, we propose that eIF5A/EF-P is a universally conserved translation elongation factor.

Your institute does not have access to this article

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Figure 1: eIF5A depletion impairs yeast cell growth and protein synthesis, and causes retention of polysomes.
Figure 2: Translation elongation defect in temperature-sensitive tif51a(D63V) mutant.
Figure 3: eIF5A stimulates elongation and termination in vitro.
Figure 4: Functional connection between eIF5A and eEF2.


  1. Merrick, W. C. & Nyborg, J. in Translational Control of Gene Expression (eds Sonenberg, N., Hershey, J. W. B. & Mathews, M. B.) 89–125 (Cold Spring Harbor Laboratory Press, 2000)

    Google Scholar 

  2. Wolff, E. C., Kang, K. R., Kim, Y. S. & Park, M. H. Posttranslational synthesis of hypusine: evolutionary progression and specificity of the hypusine modification. Amino Acids 33, 341–350 (2007)

    CAS  Article  Google Scholar 

  3. Kemper, W. M., Berry, K. W. & Merrick, W. C. Purification and properties of rabbit reticulocyte protein synthesis initiation factors M2Bα and M2Bβ. J. Biol. Chem. 251, 5551–5557 (1976)

    CAS  PubMed  Google Scholar 

  4. Schreier, M. H., Erni, B. & Staehelin, T. Initiation of mammalian protein synthesis: purification and characterization of seven initiation factors. J. Mol. Biol. 116, 727–753 (1977)

    CAS  Article  Google Scholar 

  5. Jao, D. L. & Chen, K. Y. Tandem affinity purification revealed the hypusine-dependent binding of eukaryotic initiation factor 5A to the translating 80S ribosomal complex. J. Cell. Biochem. 97, 583–598 (2006)

    CAS  Article  Google Scholar 

  6. Zanelli, C. F. et al. eIF5A binds to translational machinery components and affects translation in yeast. Biochem. Biophys. Res. Commun. 348, 1358–1366 (2006)

    CAS  Article  Google Scholar 

  7. Park, M. H., Wolff, E. C., Smit-McBride, Z., Hershey, J. W. & Folk, J. E. Comparison of the activities of variant forms of eIF-4D. The requirement for hypusine or deoxyhypusine. J. Biol. Chem. 266, 7988–7994 (1991)

    CAS  PubMed  Google Scholar 

  8. Benne, R. & Hershey, J. W. B. The mechanism of action of protein synthesis initiation factors from rabbit reticulocytes. J. Biol. Chem. 253, 3078–3087 (1978)

    CAS  PubMed  Google Scholar 

  9. Kang, H. A. & Hershey, J. W. Effect of initiation factor eIF-5A depletion on protein synthesis and proliferation of Saccharomyces cerevisiae . J. Biol. Chem. 269, 3934–3940 (1994)

    CAS  PubMed  Google Scholar 

  10. Schrader, R., Young, C., Kozian, D., Hoffmann, R. & Lottspeich, F. Temperature-sensitive eIF5A mutant accumulates transcripts targeted to the nonsense-mediated decay pathway. J. Biol. Chem. 281, 35336–35346 (2006)

    CAS  Article  Google Scholar 

  11. Zuk, D. & Jacobson, A. A single amino acid substitution in yeast eIF-5A results in mRNA stabilization. EMBO J. 17, 2914–2925 (1998)

    CAS  Article  Google Scholar 

  12. Ruhl, M. et al. Eukaryotic initiation factor 5A is a cellular target of the human immunodeficiency virus type 1 Rev activation domain mediating trans-activation. J. Cell Biol. 123, 1309–1320 (1993)

    CAS  Article  Google Scholar 

  13. Zanelli, C. F. & Valentini, S. R. Is there a role for eIF5A in translation? Amino Acids 33, 351–358 (2007)

    CAS  Article  Google Scholar 

  14. Hanawa-Suetsugu, K. et al. Crystal structure of elongation factor P from Thermus thermophilus HB8. Proc. Natl Acad. Sci. USA 101, 9595–9600 (2004)

    ADS  Article  Google Scholar 

  15. Kyrpides, N. C. & Woese, C. R. Universally conserved translation initiation factors. Proc. Natl Acad. Sci. USA 95, 224–228 (1998)

    ADS  CAS  Article  Google Scholar 

  16. Jivotovskaya, A. V., Valasek, L., Hinnebusch, A. G. & Nielsen, K. H. Eukaryotic translation initiation factor 3 (eIF3) and eIF2 can promote mRNA binding to 40S subunits independently of eIF4G in yeast. Mol. Cell. Biol. 26, 1355–1372 (2006)

    CAS  Article  Google Scholar 

  17. Smirnov, V. N. et al. Recessive nonsense-suppression in yeast: further characterization of a defect in translation. FEBS Lett. 66, 12–15 (1976)

    CAS  Article  Google Scholar 

  18. Anand, M., Chakraburtty, K., Marton, M. J., Hinnebusch, A. G. & Kinzy, T. G. Functional interactions between yeast translation eukaryotic elongation factor (eEF) 1A and eEF3. J. Biol. Chem. 278, 6985–6991 (2003)

    CAS  Article  Google Scholar 

  19. Ortiz, P. A. & Kinzy, T. G. Dominant-negative mutant phenotypes and the regulation of translation elongation factor 2 levels in yeast. Nucleic Acids Res. 33, 5740–5748 (2005)

    CAS  Article  Google Scholar 

  20. Fan, H. & Penman, S. Regulation of protein synthesis in mammalian cells. II. Inhibition of protein synthesis at the level of initiation during mitosis. J. Mol. Biol. 50, 655–670 (1970)

    CAS  Article  Google Scholar 

  21. Nielsen, P. J. & McConkey, E. H. Evidence for control of protein synthesis in HeLa cells via the elongation rate. J. Cell. Physiol. 104, 269–281 (1980)

    CAS  Article  Google Scholar 

  22. Acker, M. G., Kolitz, S. E., Mitchell, S. F., Nanda, J. S. & Lorsch, J. R. Reconstitution of yeast translation initiation. Methods Enzymol. 430, 111–145 (2007)

    CAS  Article  Google Scholar 

  23. Youngman, E. M., McDonald, M. E. & Green, R. Peptide release on the ribosome: mechanism and implications for translational control. Annu. Rev. Microbiol. 62, 353–373 (2008)

    CAS  Article  Google Scholar 

  24. Harger, J. W., Meskauskas, A., Nielsen, J., Justice, M. C. & Dinman, J. D. Ty1 retrotransposition and programmed +1 ribosomal frameshifting require the integrity of the protein synthetic translocation step. Virology 286, 216–224 (2001)

    CAS  Article  Google Scholar 

  25. Zhang, S. et al. Polysome-associated mRNAs are substrates for the nonsense-mediated mRNA decay pathway in Saccharomyces cerevisiae . RNA 3, 234–244 (1997)

    CAS  PubMed  PubMed Central  Google Scholar 

  26. Glick, B. R. & Ganoza, M. C. Identification of a soluble protein that stimulates peptide bond synthesis. Proc. Natl Acad. Sci. USA 72, 4257–4260 (1975)

    ADS  CAS  Article  Google Scholar 

  27. Ganoza, M. C. & Aoki, H. Peptide bond synthesis: function of the efp gene product. Biol. Chem. 381, 553–559 (2000)

    CAS  Article  Google Scholar 

  28. Dohmen, R. J., Wu, P. & Varshavsky, A. Heat-inducible degron: a method for constructing temperature-sensitive mutants. Science 263, 1273–1276 (1994)

    ADS  CAS  Article  Google Scholar 

  29. Labib, K., Tercero, J. A. & Diffley, J. F. Uninterrupted MCM2–7 function required for DNA replication fork progression. Science 288, 1643–1647 (2000)

    ADS  CAS  Article  Google Scholar 

  30. Harger, J. W. & Dinman, J. D. An in vivo dual-luciferase assay system for studying translational recoding in the yeast Saccharomyces cerevisiae . RNA 9, 1019–1024 (2003)

    CAS  Article  Google Scholar 

  31. Gietz, R. D. & Sugino, A. New yeast–Escherichia coli shuttle vectors constructed with in vitro mutagenized yeast genes lacking six-base pair restriction sites. Gene 74, 527–534 (1988)

    CAS  Article  Google Scholar 

  32. Tong, A. H. et al. Systematic genetic analysis with ordered arrays of yeast deletion mutants. Science 294, 2364–2368 (2001)

    ADS  CAS  Article  Google Scholar 

  33. Shenton, D. et al. Global translational responses to oxidative stress impact upon multiple levels of protein synthesis. J. Biol. Chem. 281, 29011–29021 (2006)

    CAS  Article  Google Scholar 

  34. Tarun, S. Z. & Sachs, A. B. A common function for mRNA 5′ and 3′ ends in translation initiation in yeast. Genes Dev. 9, 2997–3007 (1995)

    CAS  Article  Google Scholar 

  35. Gallie, D. R., Feder, J. N., Schimke, R. T. & Walbot, V. Post-transcriptional regulation in higher eukaryotes: the role of the reporter gene in controlling expression. Mol. Gen. Genet. 228, 258–264 (1991)

    CAS  Article  Google Scholar 

Download references


We thank T. G. Kinzy for providing anti-yeast eEF1A, eEF2 and eEF3 antisera, as well as constructs for the purification of eEF2 and eEF3; J. Lorsch for providing constructs for purification of yeast initiation factors; J. Dinmann for frameshifting reporter vectors, and A. Hinnebusch, T. G. Kinzy, A. Jivotovskaya, D. Shelton, C. Grant, J. Lorsch and members of the Dever and Hinnebusch laboratories for comments and discussion. Salary support provided by HHMI (R.G.) and NIH (D.E.E.). This work was supported in part by the Intramural Research Program of the NIH, NICHD (to T.E.D.).

Author Contributions P.S., D.E.E., R.G. and T.E.D. designed the experiments and wrote the manuscript. P.S. performed all experiments except reconstituted translation experiments, which were performed by D.E.E.

Author information

Authors and Affiliations


Corresponding author

Correspondence to Thomas E. Dever.

Supplementary information

Supplementary Information

This file contains Supplementary Tables 1-2, Supplementary Figures 1-5 with Legends and Supplementary References. (PDF 8689 kb)

PowerPoint slides

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Saini, P., Eyler, D., Green, R. et al. Hypusine-containing protein eIF5A promotes translation elongation. Nature 459, 118–121 (2009).

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI:

Further reading


By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.


Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing