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Effects of novel C-methylated spermidine analogs on cell growth via hypusination of eukaryotic translation initiation factor 5A

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Abstract

The polyamines, putrescine, spermidine, and spermine, are ubiquitous multifunctional cations essential for cellular proliferation. One specific function of spermidine in cell growth is its role as a butylamine donor for hypusine synthesis in the eukaryotic initiation factor 5A (eIF5A). Here, we report the ability of novel mono-methylated spermidine analogs (α-MeSpd, β-MeSpd, γ-MeSpd, and ω-MeSpd) to function in the hypusination of eIF5A and in supporting the growth of DFMO-treated DU145 cells. We also tested them as substrates and inhibitors for deoxyhypusine synthase (DHS) in vitro. Of these compounds, α-MeSpd, β-MeSpd, and γ-MeSpd (but not ω-MeSpd) were substrates for DHS in vitro, while they all inhibited the enzyme reaction. As racemic mixtures, only α-MeSpd and β-MeSpd supported long-term growth (9–18 days) of spermidine-depleted DU145 cells, whereas γ-MeSpd and ω-MeSpd did not. The S-enantiomer of α-MeSpd, which supported long-term growth, was a good substrate for DHS in vitro, whereas the R-isomer was not. The long-term growth of DFMO-treated cells correlated with the hypusine modification of eIF5A by intracellular methylated spermidine analogs. These results underscore the critical requirement for hypusine modification in mammalian cell proliferation and provide new insights into the specificity of the deoxyhypusine synthase reaction.

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Abbreviations

eIF5A:

Eukaryotic translation initiation factor 5A

DHS:

Deoxyhypusine synthase

DOHH:

Deoxyhypusine hydroxylase

DFMO:

α-Difluoromethylornithine

DTT:

Dithiothreitol

AG:

Aminoguanidine

Spd:

Spermidine

Spm:

Spermine

Put:

Putrescine

MeSpd:

Methylated spermidine

MeSpm:

Methylated spermine

SSAT1:

Spermidine/spermine N1-acetyltransferase

APAO:

Acetylpolyamine oxidase

TCA:

Trichloroacetic acid

DENSpm:

N 1,N 11-Diethylnorspermine

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Acknowledgments

The research was supported in part by the Intramural Research Program of National Institute of Dental and Craniofacial Research (NIDCR), NIH and by grants from the Academy of Finland, the Russian Foundation for Basic Research [grant number 09-04-01272], the program Molecular and Cell Biology of the Presidium of the Russian Academy of Sciences, the North Savo Regional Fund of the Finnish Cultural Foundation and by the University of Eastern Finland (Novel Materials, Technologies and Drug Molecules Based on Phosphorus and/or Nitrogen Compounds; JV/TAK). The authors thank Ms. Sisko Juutinen, Ms. Anne Karppinen, Ms Arja Korhonen, and Ms. Tuula Reponen for skillful technical assistance and Dr. Edith C. Wolff (NIDCR, NIH) for critical reading of the manuscript and helpful suggestions.

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Authors and Affiliations

  1. A.I. Virtanen Institute for Molecular Sciences, Biocenter Kuopio, University of Eastern Finland, Yliopistonranta 1E, 70210, Kuopio, Finland

    Mervi T. Hyvönen & Leena Alhonen

  2. Department of Biosciences, Laboratory of Chemistry, Biocenter Kuopio, University of Eastern Finland, Yliopistonranta 1E, 70210, Kuopio, Finland

    Tuomo A. Keinänen & Jouko Vepsäläinen

  3. Oral and Pharyngeal Cancer Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD, 20892, USA

    Jong Hwan Park & Myung Hee Park

  4. Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov Street 32, Moscow, 119991, Russia

    Maxim Khomutov, Alina Simonian & Alex R. Khomutov

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  1. Mervi T. Hyvönen
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  2. Tuomo A. Keinänen
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  3. Maxim Khomutov
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  5. Jouko Vepsäläinen
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  6. Jong Hwan Park
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  7. Alex R. Khomutov
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  8. Leena Alhonen
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  9. Myung Hee Park
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Correspondence to Mervi T. Hyvönen or Myung Hee Park.

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Hyvönen, M.T., Keinänen, T.A., Khomutov, M. et al. Effects of novel C-methylated spermidine analogs on cell growth via hypusination of eukaryotic translation initiation factor 5A. Amino Acids 42, 685–695 (2012). https://doi.org/10.1007/s00726-011-0984-1

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