Arabidopsis CYP735A1 and CYP735A2 encode cytokinin hydroxylases that catalyze the biosynthesis of trans-Zeatin.
Cytokinins (CKs), a group of phytohormones, are adenine derivatives that carry either an isoprene-derived or an aromatic side chain at the N(6) terminus. trans-Zeatin (tZ), an isoprenoid CK, is assumed to play a central physiological role because of its general occurrence and high activity in bioassays. Although hydroxylation of isopentenyladenine-type CKs is a key step of tZ biosynthesis, the catalyzing enzyme has not been characterized yet. Here we demonstrate that CYP735A1 and CYP735A2 are cytochrome P450 monooxygenases (P450s) that catalyze the biosynthesis of tZ. We identified the genes from Arabidopsis using an adenosine phosphate-isopentenyltransferase (AtIPT4)/P450 co-expression system in yeast. Co-expression of AtIPT4 and CYP735A enabled yeast to excrete tZ and the nucleosides to the culture medium. In vitro, both CYP735As preferentially utilized isopentenyladenine nucleotides rather than the nucleoside and free base forms and produced tZ nucleotides but not the cis-isomer. The expression of CYP735A1 and CYP735A2 was differentially regulated in terms of organ specificity and response to CK. Root-specific induction of CYP735A2 expression by CK suggests that the trans-hydroxylation is involved in the regulation of CK metabolism and signaling in roots.
Plant Science Center, RIKEN, Suehiro 1-7-22, Tsurumi, Yokohama 230-0045, Japan.
Kentaro
K
Takei
Tomoyuki
T
Yamaya
Hitoshi
H
Sakakibara
D029681:Arabidopsis Proteins; D002384:Catalysis; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D003583:Cytokinins / Q000378:metabolism*; D018506:Gene Expression Regulation, Plant; D006900:Hydroxylation; D006899:Mixed Function Oxygenases / Q000235:genetics / Q000378:metabolism*; D010937:Plant Growth Regulators / Q000378:metabolism; D018517:Plant Roots / Q000378:metabolism; D018514:Plant Structures / Q000378:metabolism; D015026:Zeatin / Q000378:metabolism*
D029681:Arabidopsis Proteins; D003583:Cytokinins; D010937:Plant Growth Regulators; D015026:Zeatin; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; C492981:cytochrome P-450 735A1, Arabidopsis; C492982:cytochrome P-450 735A2, Arabidopsis