Independent roles of eIF5A and polyamines in cell proliferation

Kazuhiro Nishimura

Kaori Murozumi

Akira Shirahata

Myung Park

Keiko Kashiwagi

Kazuei Igarashi

*Graduate School of Pharmaceutical Sciences, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba 260-8675, Japan

†Faculty of Pharmaceutical Sciences, Josai University, 1-1 Keyakidai, Sakado 350-0248, Japan

‡Oral and Pharyngeal Cancer Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD 20892-4340, U.S.A.

^{To whom correspondence should be addressed (email}pj.ca.u-abihc.p@77061agi).

Received 2004 Aug 31; Revised 2004 Sep 16; Accepted 2004 Sep 17.

Abstract

To examine the roles of active hypusinated eIF5A (eukaryotic translation initiation factor 5A) and polyamines in cell proliferation, mouse mammary carcinoma FM3A cells were treated with an inhibitor of deoxyhypusine synthase, GC₇ (N^{-guanyl-1, 7-diaminoheptane), or with an inhibitor of ornithine decarboxylase, DFMO (α-difluoromethylornithine), or with DFMO plus an inhibitor of spermine synthase, APCHA [}N^{-(3-aminopropyl)-cyclohexylamine]. Treatment with GC}₇ decreased the level of active eIF5A on day 1 without affecting cellular polyamine content, and inhibition of cell growth occurred from day 2. This delay reflects the fact that eIF5A was present in excess and was very stable in these cells. Treatment with DFMO or with DFMO plus APCHA inhibited cell growth on day 1. DFMO considerably decreased the levels of putrescine and spermidine, and the formation of active eIF5A began to decrease when the level of spermidine fell below 8 nmol/mg of protein after 12 h of incubation with DFMO. The combination of DFMO and APCHA markedly decreased the levels of putrescine and spermine and significantly decreased the level of spermidine, but did not affect the level of active eIF5A until day 3 when spermidine level decreased to 7 nmol/mg of protein. The results show that a decrease in either active eIF5A or polyamines inhibits cell growth, indicating that eIF5A and polyamines are independently involved in cell growth.

Keywords: biosynthetic inhibitor, cell proliferation, deoxyhypusine synthase, eukaryotic translation initiation factor 5A (eIF5A), hypusine, polyamine

Abbreviations: APCHA, N^{-(3-aminopropyl)cyclohexylamine; DFMO, α-difluoromethylornithine; eIF5A, eukaryotic translation initiation factor 5A; FUT-175, 6-amino-2-naphthyl-4-guanidinobenzoate dihydrochloride; GC}₇, N^{-guanyl-1,7-diaminoheptane}

Abstract

Acknowledgments

We thank Dr K. Williams and Dr A. J. Michael for their help in preparing this manuscript. We also thank Aventis Pharma and Torii Pharmaceutical for providing DFMO and FUT-175. This work was supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology, Japan.

Acknowledgments

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