Amino Acids

, Volume 42, Issue 2, pp 867–876

Constitutively and highly expressed Oryza sativa polyamine oxidases localize in peroxisomes and catalyze polyamine back conversion

  • Yusuke Ono
  • Dong Wook Kim
  • Kanako Watanabe
  • Ayano Sasaki
  • Masaru Niitsu
  • Thomas Berberich
  • Tomonobu Kusano
  • Yoshihiro Takahashi
Original Article

DOI: 10.1007/s00726-011-1002-3

Cite this article as:
Ono, Y., Kim, D.W., Watanabe, K. et al. Amino Acids (2012) 42: 867. doi:10.1007/s00726-011-1002-3

Abstract

Polyamine oxidases (PAOs) are FAD-dependent enzymes involved in polyamine (PA) catabolism. Recent studies have revealed that plant PAOs are not only active in the terminal catabolism of PAs as demonstrated for maize apoplastic PAO but also in a polyamine back-conversion pathway as shown for most Arabidopsis PAOs. We have characterized Oryza sativaPAOs at molecular and biochemical levels. The rice genome contains 7 PAO isoforms that are termed OsPAO1 to OsPAO7. Of the seven PAOs, OsPAO3, OsPAO4, and OsPAO5 transcripts were most abundant in 2-week-old seedlings and mature plants, while OsPAO1, OsPAO2, OsPAO6, and OsPAO7 were expressed at very low levels with different tissue specificities. The more abundantly expressed PAOs—OsPAO3, OsPAO4, and OsPAO5—were cloned, and their gene products were produced in Escherichia coli. The enzymatic activities of the purified OsPAO3 to OsPAO5 proteins were examined. OsPAO3 favored spermidine (Spd) as substrate followed by thermospermine (T-Spm) and spermine (Spm) and showed a full PA back-conversion activity. OsPAO4 substrate specificity was similar to that of OsPAO5 preferring Spm and T-Spm but not Spd. Those enzymes also converted Spm and T-Spm to Spd, again indicative of PA back-conversion activities. Lastly, we show that OsPAO3, OsPAO4, and OsPAO5 are localized in peroxisomes. Together, these data revealed that constitutively and highly expressed O. sativa PAOs are localized in peroxisomes and catalyze PA back-conversion processes.

Keywords

Oryza sativa Peroxisome Polyamine Polyamine oxidase Substrate specificity Tissue specificity 

Abbreviations

AO

Amine oxidase

Arg

Arginine

AtPAO

Arabidopsis thaliana polyamine oxidase

DAS

Day(s) after sowing

FAD

Flavin adenine dinucleotide

GFP

Green fluorescent protein

HvPAO

Barley polyamine oxidase

NorSpm

Norspermine

N1-AcSpm

N1-acetylspermine

Orn

Ornithine

PA

Polyamine

PAO

Polyamine oxidase

PCR

Polymerase chain reaction

OsPAO

Oryza sativa polyamine oxidase

PTS

Peroxisomal targeting signal

Put

Putrescine

Spd

Spermidine

Spm

Spermine

T-Spm

Thermospermine

ZmPAO

Maize polyamine oxidase

Supplementary material

726_2011_1002_MOESM1_ESM.doc (36 kb)
Supplementary material 1 (DOC 36 kb)

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Yusuke Ono
    • 1
  • Dong Wook Kim
    • 1
  • Kanako Watanabe
    • 1
  • Ayano Sasaki
    • 1
  • Masaru Niitsu
    • 2
  • Thomas Berberich
    • 3
  • Tomonobu Kusano
    • 1
  • Yoshihiro Takahashi
    • 1
  1. 1.Graduate School of Life SciencesTohoku UniversitySendai, MiyagiJapan
  2. 2.Faculty of Pharmaceutical Sciences, Josai UniversitySakado, SaitamaJapan
  3. 3.Biodiversity and Climate Research Center FrankfurtFrankfurtGermany