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A maize spermine synthase 1 PEST sequence fused to the GUS reporter protein facilitates proteolytic degradation
26S proteasome; Aminopropyl transferases; PEST; Polyamines; Spermine synthase ATP dependent 26S protease; benzyloxycarbonylleucyl-leucyl-leucine aldehyde; beta glucuronidase; cysteine proteinase inhibitor; hybrid protein; leupeptin; proteasome; spermine synthase; vegetable protein; amino acid sequence; Arabidopsis; cytology; genetics; maize; metabolism; molecular genetics; onion; plant gene; protein degradation; reverse transcription polymerase chain reaction; transgenic plant; Western blotting; Amino Acid Sequence; Arabidopsis; Blotting, Western; Cysteine Proteinase Inhibitors; Genes, Plant; Glucuronidase; Leupeptins; Molecular Sequence Data; Onions; Plant Proteins; Plants, Genetically Modified; Proteasome Endopeptidase Complex; Proteolysis; Recombinant Fusion Proteins; Reverse Transcriptase Polymerase Chain Reaction; Spermine Synthase; Zea mays
beta glucuronidase, 9001-45-0; leupeptin, 54577-99-0; proteasome, 140879-24-9; spermine synthase, 74812-43-4; ATP dependent 26S protease; benzyloxycarbonylleucyl-leucyl-leucine aldehyde; Cysteine Proteinase Inhibitors; Glucuronidase; Leupeptins; Plant Proteins; Proteasome Endopeptidase Complex; Recombinant Fusion Proteins; Spermine Synthase
Polyamines are low molecular weight aliphatic compounds involved in various biochemical, cellular and physiological processes in all organisms. In plants, genes involved in polyamine biosynthesis and catabolism are regulated at transcriptional, translational, and posttranslational level. In this research, we focused on the characterization of a PEST sequence (rich in proline, glutamic acid, serine, and threonine) of the maize spermine synthase 1 (ZmSPMS1). To this aim, 123 bp encoding 40 amino acids of the C-terminal region of the ZmSPMS1 enzyme containing the PEST sequence were fused to the GUS reporter gene. This fusion was evaluated in Arabidopsis thaliana transgenic lines and onion monolayers transient expression system. The ZmSPMS1 PEST sequence leads to specific degradation of the GUS reporter protein. It is suggested that the 26S proteasome may be involved in GUS::PEST fusion degradation in both onion and Arabidopsis. The PEST sequences appear to be present in plant spermine synthases, mainly in monocots. © 2014 Elsevier Masson SAS.
Consejo Nacional de Ciencia y Tecnología, CONACYT: 2008–103106
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