EIF5A Functions Globally in Translation Elongation and Termination

Overview

Journal Mol Cell

Publisher Cell Press

Specialty Cell Biology

Date 2017 Apr 11

PMID 28392174

Citations 235

Authors

Anthony P Schuller

Colin Chih-Chien Wu

Thomas E Dever

Allen R Buskirk

Rachel Green

Affiliations

Soon will be listed here.

Abstract

The eukaryotic translation factor eIF5A, originally identified as an initiation factor, was later shown to promote translation elongation of iterated proline sequences. Using a combination of ribosome profiling and in vitro biochemistry, we report a much broader role for eIF5A in elongation and uncover a critical function for eIF5A in termination. Ribosome profiling of an eIF5A-depleted strain reveals a global elongation defect, with abundant ribosomes stalling at many sequences, not limited to proline stretches. Our data also show ribosome accumulation at stop codons and in the 3' UTR, suggesting a global defect in termination in the absence of eIF5A. Using an in vitro reconstituted translation system, we find that eIF5A strongly promotes the translation of the stalling sequences identified by profiling and increases the rate of peptidyl-tRNA hydrolysis more than 17-fold. We conclude that eIF5A functions broadly in elongation and termination, rationalizing its high cellular abundance and essential nature.

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References

Blaha G, Stanley R, Steitz T . Formation of the first peptide bond: the structure of EF-P bound to the 70S ribosome. Science. 2009; 325(5943):966-70. PMC: 3296453. DOI: 10.1126/science.1175800. View

Acker M, Kolitz S, Mitchell S, Nanda J, Lorsch J . Reconstitution of yeast translation initiation. Methods Enzymol. 2007; 430:111-45. DOI: 10.1016/S0076-6879(07)30006-2. View

Katoh T, Wohlgemuth I, Nagano M, Rodnina M, Suga H . Essential structural elements in tRNA(Pro) for EF-P-mediated alleviation of translation stalling. Nat Commun. 2016; 7:11657. PMC: 4890201. DOI: 10.1038/ncomms11657. View

Ude S, Lassak J, Starosta A, Kraxenberger T, Wilson D, Jung K . Translation elongation factor EF-P alleviates ribosome stalling at polyproline stretches. Science. 2012; 339(6115):82-5. DOI: 10.1126/science.1228985. View

Ingolia N, Ghaemmaghami S, Newman J, Weissman J . Genome-wide analysis in vivo of translation with nucleotide resolution using ribosome profiling. Science. 2009; 324(5924):218-23. PMC: 2746483. DOI: 10.1126/science.1168978. View

Nishimura K, Kanemaki M . Rapid Depletion of Budding Yeast Proteins via the Fusion of an Auxin-Inducible Degron (AID). Curr Protoc Cell Biol. 2014; 64:20.9.1-16. DOI: 10.1002/0471143030.cb2009s64. View

Gregio A, Cano V, Avaca J, Valentini S, Zanelli C . eIF5A has a function in the elongation step of translation in yeast. Biochem Biophys Res Commun. 2009; 380(4):785-90. DOI: 10.1016/j.bbrc.2009.01.148. View

Brown A, Shao S, Murray J, Hegde R, Ramakrishnan V . Structural basis for stop codon recognition in eukaryotes. Nature. 2015; 524(7566):493-496. PMC: 4591471. DOI: 10.1038/nature14896. View

Brown C, Stockwell P, Trotman C, Tate W . Sequence analysis suggests that tetra-nucleotides signal the termination of protein synthesis in eukaryotes. Nucleic Acids Res. 1990; 18(21):6339-45. PMC: 332501. DOI: 10.1093/nar/18.21.6339. View

10.

SCHREIER M, Erni B, Staehelin T . Initiation of mammalian protein synthesis. I. Purification and characterization of seven initiation factors. J Mol Biol. 1977; 116(4):727-53. DOI: 10.1016/0022-2836(77)90268-6. View

11.

Eyler D, Green R . Distinct response of yeast ribosomes to a miscoding event during translation. RNA. 2011; 17(5):925-32. PMC: 3078741. DOI: 10.1261/rna.2623711. View

12.

Pierson W, Hoffer E, Keedy H, Simms C, Dunham C, Zaher H . Uniformity of Peptide Release Is Maintained by Methylation of Release Factors. Cell Rep. 2016; 17(1):11-18. PMC: 5079439. DOI: 10.1016/j.celrep.2016.08.085. View

13.

Benne R, Hershey J . The mechanism of action of protein synthesis initiation factors from rabbit reticulocytes. J Biol Chem. 1978; 253(9):3078-87. View

14.

Sabi R, Tuller T . Modelling the efficiency of codon-tRNA interactions based on codon usage bias. DNA Res. 2014; 21(5):511-26. PMC: 4195497. DOI: 10.1093/dnares/dsu017. View

15.

Guydosh N, Green R . Dom34 rescues ribosomes in 3' untranslated regions. Cell. 2014; 156(5):950-62. PMC: 4022138. DOI: 10.1016/j.cell.2014.02.006. View

16.

Saini P, Eyler D, Green R, Dever T . Hypusine-containing protein eIF5A promotes translation elongation. Nature. 2009; 459(7243):118-21. PMC: 3140696. DOI: 10.1038/nature08034. View

17.

Glick B, Ganoza M . Identification of a soluble protein that stimulates peptide bond synthesis. Proc Natl Acad Sci U S A. 1975; 72(11):4257-60. PMC: 388699. DOI: 10.1073/pnas.72.11.4257. View

18.

Woolstenhulme C, Guydosh N, Green R, Buskirk A . High-precision analysis of translational pausing by ribosome profiling in bacteria lacking EFP. Cell Rep. 2015; 11(1):13-21. PMC: 4835038. DOI: 10.1016/j.celrep.2015.03.014. View

19.

Park M, Cooper H, Folk J . Identification of hypusine, an unusual amino acid, in a protein from human lymphocytes and of spermidine as its biosynthetic precursor. Proc Natl Acad Sci U S A. 1981; 78(5):2869-73. PMC: 319460. DOI: 10.1073/pnas.78.5.2869. View

20.

Rossi D, Barbosa N, Galvao F, Boldrin P, Hershey J, Zanelli C . Evidence for a Negative Cooperativity between eIF5A and eEF2 on Binding to the Ribosome. PLoS One. 2016; 11(4):e0154205. PMC: 4845985. DOI: 10.1371/journal.pone.0154205. View