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Originally published In Press as doi:10.1074/jbc.M008544200 on February 14, 2001

J. Biol. Chem., Vol. 276, Issue 20, 17541-17549, May 18, 2001
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Isolation and Characterization of Senescence-induced cDNAs Encoding Deoxyhypusine Synthase and Eucaryotic Translation Initiation Factor 5A from Tomato*

Tzann-Wei Wang, Lily Lu, Denis Wang, and John E. ThompsonDagger

From the Department of Biology, University of Waterloo, Waterloo, Ontario N2L 3G1, Canada

Full-length cDNA clones encoding deoxyhypusine synthase (DHS) and eucaryotic initiation factor 5A (eIF-5A) have been isolated from a cDNA expression library prepared from tomato leaves (Lycopersicon esculentum, cv. Match) exposed to environmental stress. DHS mediates the first of two enzymatic reactions that activate eIF-5A by converting a conserved lysine to the unusual amino acid, deoxyhypusine. Recombinant protein obtained by expressing tomato DHS cDNA in Escherichia coli proved capable of carrying out the deoxyhypusine synthase reaction in vitro in the presence of eIF-5A. Of particular interest is the finding that DHS mRNA and eIF-5A mRNA show a parallel increase in abundance in senescing tomato flowers, senescing tomato fruit, and environmentally stressed tomato leaves exhibiting programmed cell death. Western blot analyses indicated that DHS protein also increases at the onset of senescence. It is apparent from previous studies with yeast and mammalian cells that hypusine-modified eIF-5A facilitates the translation of a subset of mRNAs mediating cell division. The present study provides evidence for senescence-induced DHS and eIF-5A in tomato tissues that may facilitate the translation of mRNA species required for programmed cell death.

* This work was supported by the Natural Sciences and Engineering Research Council of Canada.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AF296077 (tomato DHS), AF296078 (Arabidopsis DHS), AF296079 (banana DHS), AF296080 (carnation DHS), AF296081 (carnation eIF-5A), AF296082 (Arabidopsis eIF-5A), AF296083 (tomato eIF-5A1), AF296084 (tomato eIF-5A2), AF296085 (tomato eIF-5A3), and AF296086 (tomato eIF-5A4).

Dagger To whom correspondence should be addressed: Dept. of Biology, University of Waterloo, Waterloo, Ontario N2L 3G1, Canada. Tel.: 519-888-4465; Fax: 519-746-2543; E-mail: jet@sciborg.uwaterloo.ca.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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