title: Constitutively and highly expressed Oryza sativa polyamine oxidases localize in peroxisomes and catalyze polyamine back conversion.
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 sativa PAOs 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.
authors.affiliation: Graduate School of Life Sciences, Tohoku University, 2-1-1 Katahira, Aoba, Sendai, Miyagi, 980-8577, Japan.
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Ono
authors.forename: Dong Wook
authors.initials: DW
authors.lastname: Kim
authors.forename: Kanako
authors.initials: K
authors.lastname: Watanabe
authors.forename: Ayano
authors.initials: A
authors.lastname: Sasaki
authors.forename: Masaru
authors.initials: M
authors.lastname: Niitsu
authors.forename: Thomas
authors.initials: T
authors.lastname: Berberich
authors.forename: Tomonobu
authors.initials: T
authors.lastname: Kusano
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Takahashi
mesh_terms: D001483:Base Sequence; D015317:Biogenic Polyamines / Q000378:metabolism*; D002384:Catalysis; D017931:DNA Primers; D012275:Oryza / Q000201:enzymology*; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000378:metabolism*; D020675:Peroxisomes / Q000201:enzymology*; D016133:Polymerase Chain Reaction; D000097667:Polyamine Oxidase
chemical_list: D015317:Biogenic Polyamines; D017931:DNA Primers; D000587:Oxidoreductases Acting on CH-NH Group Donors; D000097667:Polyamine Oxidase