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Putrescine overproduction does not affect the catabolism of spermidine and spermine in poplar and Arabidopsis

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Abstract

The effect of up-regulation of putrescine (Put) production by genetic manipulation on the turnover of spermidine (Spd) and spermine (Spm) was investigated in transgenic cells of poplar (Populus nigra × maximowiczii) and seedlings of Arabidopsis thaliana. Several-fold increase in Put production was achieved by expressing a mouse ornithine decarboxylase cDNA either under the control of a constitutive (in poplar) or an inducible (in Arabidopsis) promoter. The transgenic poplar cells produced and accumulated 8–10 times higher amounts of Put than the non-transgenic cells, whereas the Arabidopsis seedlings accumulated up to 40-fold higher amounts of Put; however, in neither case the cellular Spd or Spm increased consistently. The rate of Spd and Spm catabolism and the half-life of cellular Spd and Spm were measured by pulse-chase experiments using [14C]Spd or [14C]Spm. Spermidine half-life was calculated to be about 22–32 h in poplar and 52–56 h in Arabidopsis. The half-life of cellular Spm was calculated to be approximately 24 h in Arabidopsis and 36–48 h in poplar. Both species were able to convert Spd to Spm and Put, and Spm to Spd and Put. The rates of Spd and Spm catabolism in both species were several-fold slower than those of Put, and the overproduction of Put had only a small effect on the overall rates of turnover of Spd or Spm. There was little effect on the rates of Spd to Spm conversion as well as the conversion of Spm into lower polyamines. While Spm was mainly converted back to Spd and not terminally degraded, Spd was removed from the cells largely through terminal catabolism in both species.

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Notes

  1. The enzymes commonly called polyamine oxidases, which oxidize Put, Spd, Spm, acetyl-Spd and acetyl-Spm, have been assigned different EC numbers depending upon the source or characterization of the substrate preference; the terminology sometimes appears confusing. A few examples of the confusion about nomenclature of different polyamine oxidases are cited here: Spermine oxidase (EC 1.5.3.16), http://www.brenda-enzymes.org/php/result_flat.php4?ecno=1.5.3.16; Polyamine oxidase (EC 1.5.3.3), http://www.ebi.ac.uk/intenz/query?cmd=SearchEC&ec=1.5.3.3; AtPAO1 (EC# 1.5.3.16), http://www.uniprot.org/uniprot/Q9FNA2; AtPAO2 (EC# 1.5.3.?), http://www.uniprot.org/uniprot/Q9SKX5, http://enzyme.expasy.org/EC/1.5.3; AtPAO3 (EC# 1.5.3.17), http://www.uniprot.org/uniprot/Q9LYT1; AtPAO4 (EC# 1.5.3.16), http://www.uniprot.org/uniprot/Q8H191; AtPAO5 ((EC# 1.5.3.?), http://www.uniprot.org/uniprot/Q9SU79, http://enzyme.expasy.org/EC/1.5.3; ZmPAO1 (EC# 1.5.3.14/1.5.3.15?), http://www.ncbi.nlm.nih.gov/nuccore/NM_001111636; Non-specific polyamine oxidase (EC 1.5.3.17), http://enzyme.expasy.org/EC/1.5.3.17; N8-acetylspermidine oxidase, http://www.brenda-enzymes.org/php/result_flat.php4?ecno=1.5.3.15; N(1)-acetylpolyamine oxidase (EC 1.5.3.13), http://enzyme.expasy.org/EC/1.5.3.13; N-acetylpolyamine oxidase EC 1.5.3.11 (deleted entry), http://www.chem.qmul.ac.uk/iubmb/enzyme/EC1/5/3/11.html, http://www.brenda-enzymes.org/php/result_flat.php4?ecno=1.5.3.11.

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Acknowledgments

This is a Scientific Contribution Number 2509 From the NHAES.

The authors would like to thank Stephanie Long and Ben Mayer from the USDA-Forest Service for technical help in the analysis of PAs, and Bernadette Glasheen and Suneet Bains for producing the poplar transgenic cell line as a part of their undergraduate research. L.S. would also like to thank Sarah Greenberg and Boubker Barchi for help in sample collection of Arabidopsis plants for HPLC analysis. This study was partially funded by NH-Agricultural Experiment Station, USDA Forest Service (NRS), UNH Graduate Research Enhancement Awards, Graduate School Summer TA fellowship (Raj M and L.S.) and an Edith Jones Fellowship to P.B.

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The authors declare that they have no conflict of interest.

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  1. Pratiksha Bhatnagar

    Present address: Pfizer Inc., 1 Burtt Road, Andover, MA, 01810, USA

Authors and Affiliations

  1. Department of Biological Sciences, University of New Hampshire, Durham, NH, 03824, USA

    Lin Shao & Subhash C. Minocha

  2. USDA, 308 Sturtevant Hall, Geneva, NY, 14456, USA

    Rajtilak Majumdar

  3. US Forest Service, Northern Research Station, Durham, NH, 03824, USA

    Rakesh Minocha

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  1. Lin Shao
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  2. Pratiksha Bhatnagar
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  5. Subhash C. Minocha
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Correspondence to Subhash C. Minocha.

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Shao, L., Bhatnagar, P., Majumdar, R. et al. Putrescine overproduction does not affect the catabolism of spermidine and spermine in poplar and Arabidopsis. Amino Acids 46, 743–757 (2014). https://doi.org/10.1007/s00726-013-1581-2

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