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eIF5A has a function in the cotranslational translocation of proteins into the ER

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Abstract

The putative eukaryotic translation initiation factor 5A (eIF5A) is a highly conserved and essential protein present in all organisms except bacteria. To be activated, eIF5A requires the conversion of a specific residue of lysine into hypusine. This hypusine modification occurs posttranslationally in two enzymatic steps, and the polyamine spermidine is the substrate. Despite having an essential function in translation elongation, the critical role played by eIF5A remains unclear. In addition to demonstrating genetic interactions with translation factors, eIF5A mutants genetically interact with mutations in YPT1, which encodes an essential protein involved in endoplasmic reticulum (ER)-to-Golgi vesicle transport. In this study, we investigated the correlation between the function of eIF5A in translation and secretion in yeast. The results of in vivo translocation assays and genetic interaction analyses suggest a specific role for eIF5A in the cotranslational translocation of proteins into the ER, but not in the posttranslational pathway. Additionally, we observed that a block in eIF5A activation up-regulates stress-induced chaperones, which also occurs when SRP function is lost. Finally, loss of eIF5A function affects binding of the ribosome-nascent chain complex to SRP. These results link eIF5A function in translation with a role of SRP in the cell and may help explain the dual effects of eIF5A in differential and general translation.

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Abbreviations

eIF5A:

Eukaryotic translation initiation factor 5A

TIF51A :

Gene encoding eIF5A in yeast

eEF2:

Eukaryotic translation elongation factor 2

EFT2 :

Gene encoding eEF2 in yeast

eEF1A:

Eukaryotic translation elongation factor 1A

EF-P:

Elongation factor P

SRP:

Signal recognition particle

ER:

Endoplasmic reticulum

RNC:

Ribosome-nascent chain

CPY:

Carboxypeptidase Y

DPAP B:

Dipeptidyl aminopeptidase B

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Acknowledgments

We are grateful to Colin Stirling (University of Newcastle, Newcastle, United Kingdom), Martin Pool (Faculty of Life Sciences, University of Manchester, Manchester, United Kingdom) and Davis Ng (Biological Science, National University of Singapore, Singapore) for their generous gifts of strains, plasmids, and antibodies. This work was supported by grants to S R V and C F Z from the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP), the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) and PADC from the Faculdade de Ciências Farmacêuticas, UNESP. We also thank the FAPESP for fellowships awarded to most of the authors.

Conflict of interest

We declare that we have no conflict of interest.

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Authors and Affiliations

  1. Department of Biological Sciences, School of Pharmaceutical Sciences, Univ Estadual Paulista, UNESP, Araraquara, SP, Brazil

    Danuza Rossi, Fabio Carrilho Galvão, Hermano Martins Bellato, Paulo E. G. Boldrin, Sandro Roberto Valentini & Cleslei Fernando Zanelli

  2. Departament of Molecular Genetics, Donnelly Center for Cellular and Biomolecular Research, University of Toronto, Toronto, Canada

    Fabio Carrilho Galvão & Brenda J. Andrews

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  1. Danuza Rossi
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  2. Fabio Carrilho Galvão
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  3. Hermano Martins Bellato
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  4. Paulo E. G. Boldrin
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  5. Brenda J. Andrews
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  6. Sandro Roberto Valentini
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  7. Cleslei Fernando Zanelli
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Correspondence to Cleslei Fernando Zanelli.

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Rossi, D., Galvão, F.C., Bellato, H.M. et al. eIF5A has a function in the cotranslational translocation of proteins into the ER. Amino Acids 46, 645–653 (2014). https://doi.org/10.1007/s00726-013-1618-6

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