Cloning and functional characterization of the Arabidopsis N-acetylserotonin O-methyltransferase responsible for melatonin synthesis.
The N-acetylserotonin O-methyltransferase (ASMT) gene encodes the enzyme that catalyzes the conversion of N-acetylserotonin to melatonin as the last step in melatonin biosynthesis. The first plant ASMT gene to be cloned was from rice. An orthologous gene encoding a protein with ASMT activity and only 39.7% amino acid sequence identity to the rice ASMT protein was recently isolated from apple (Malus zumi). The low homology of the apple ASMT sequence prompted us to screen the Arabidopsis genome for a homologous ASMT gene. The At4g35160 gene exhibited the highest sequence identity (31%) to the rice ASMT gene, followed by the At1g76790 gene with 29% sequence identity. We purified recombinant proteins expressed from the two Arabidopsis genes. The At4g35160 recombinant protein exhibited ASMT enzyme activity, but the At1g76790 recombinant protein did not; thus, we designated At4g35160 as an Arabidopsis thaliana ASMT (AtASMT) gene. The AtASMT protein catalyzed the conversion of N-acetylserotonin to melatonin and serotonin to 5-methoxytryptamine with Vmax values of 0.11 and 0.29 pkat/mg protein, respectively. However, AtASMT exhibited no caffeic acid O-methyltransferase activity, suggesting that its function was highly specific to melatonin synthesis. AtASMT transcripts were induced by cadmium treatment in Arabidopsis followed by increased melatonin synthesis. Similar to other ASMT proteins, AtASMT was localized in the cytoplasm and its ectopic overexpression in rice resulted in increased ASMT enzyme activity and melatonin production, indicating the involvement of AtASMT in melatonin synthesis.
Department of Biotechnology, Bioenergy Research Center, College of Agriculture and Life Sciences, Chonnam National University, Gwangju, Korea.
Yeong
Y
Byeon
Department of Biotechnology, Bioenergy Research Center, College of Agriculture and Life Sciences, Chonnam National University, Gwangju, Korea.
Hye-Jung
HJ
Lee
Department of Biotechnology, Bioenergy Research Center, College of Agriculture and Life Sciences, Chonnam National University, Gwangju, Korea.
Hyoung Yool
HY
Lee
Department of Biotechnology, Bioenergy Research Center, College of Agriculture and Life Sciences, Chonnam National University, Gwangju, Korea.
Kyoungwhan
K
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D000121:Acetylserotonin O-Methyltransferase / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics; D029681:Arabidopsis Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D003001:Cloning, Molecular; D008550:Melatonin / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D012275:Oryza / Q000201:enzymology / Q000235:genetics; D030821:Plants, Genetically Modified / Q000201:enzymology / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics
D029681:Arabidopsis Proteins; D011994:Recombinant Proteins; D000121:Acetylserotonin O-Methyltransferase; D008550:Melatonin
Arabidopsis O-methyltransferases; N-acetylserotonin O-methyltransferase; melatonin; rice ASMT ortholog; transgenic rice