Abrogation of upstream open reading frame-mediated translational control of a plant S-adenosylmethionine decarboxylase results in polyamine disruption and growth perturbations.
S-Adenosylmethionine decarboxylase (AdoMetDC) is a key enzyme in polyamine biosynthesis. We show that the plant AdoMetDC activity is subject to post-transcriptional control by polyamines. A highly conserved small upstream open reading frame (uORF) in the AdoMetDC mRNA 5' leader is responsible for translational repression of a downstream beta-glucuronidase reporter cistron in transgenic tobacco plants. Elimination of the small uORF from an AdoMetDC cDNA led to increased relative translational efficiency of the AdoMetDC proenzyme in transgenic plants. The resulting increased activity of AdoMetDC caused disruption to polyamine levels with depletion of putrescine, reduction of spermine levels, and a more than 400-fold increase in the level of decarboxylated S-adenosylmethionine. These changes were associated with severe growth and developmental defects. The high level of decarboxylated S-adenosylmethionine was not associated with any change in 5'-methylcytosine content in genomic DNA and S-adenosylmethionine levels were more or less normal, indicating a highly efficient system for maintenance of S-adenosylmethionine levels in plants. This work demonstrates that uORF-mediated translational control of AdoMetDC is essential for polyamine homeostasis and for normal growth and development.
Hanfrey
Colin
C
Division of Food Safety Science, Institute of Food Research, Norwich Research Park, Colney, United Kingdom.
Franceschetti
Marina
M
Mayer
Melinda J
MJ
Illingworth
Crista
C
Michael
Anthony J
AJ
D012437:Adenosylmethionine Decarboxylase / Q000096:biosynthesis* / Q000235:genetics*; D001483:Base Sequence; D002455:Cell Division; D002851:Chromatography, High Pressure Liquid; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D016366:Open Reading Frames; D018515:Plant Leaves / Q000378:metabolism; D030821:Plants, Genetically Modified; D010957:Plasmids / Q000378:metabolism; D011073:Polyamines / Q000378:metabolism*; D014176:Protein Biosynthesis; D012313:RNA / Q000378:metabolism; D012323:RNA Processing, Post-Transcriptional; D012333:RNA, Messenger / Q000378:metabolism; D014026:Nicotiana / Q000235:genetics; D014158:Transcription, Genetic
D011073:Polyamines; D012333:RNA, Messenger; D012313:RNA; D012437:Adenosylmethionine Decarboxylase