A polyamine metabolon involving aminopropyl transferase complexes in Arabidopsis.
The conversion of putrescine to spermidine in the biosynthetic pathway of plant polyamines is catalyzed by two closely related spermidine synthases, SPDS1 and SPDS2, in Arabidopsis. In the yeast two-hybrid system, SPDS2 was found to interact with SPDS1 and a novel protein, SPMS (spermine synthase), which is homologous with SPDS2 and SPDS1. SPMS interacts with both SPDS1 and SPDS2 in yeast and in vitro. Unlike SPDS1 and SPDS2, SPMS failed to suppress the speDelta3 deficiency of spermidine synthase in yeast. However, SPMS was able to complement the speDelta4 spermine deficiency in yeast, indicating that SPMS is a novel spermine synthase. The SPDS and SPMS proteins showed no homodimerization but formed heterodimers in vitro. Pairwise coexpression of hemagglutinin- and c-Myc epitope-labeled proteins in Arabidopsis cells confirmed the existence of coimmunoprecipitating SPDS1-SPDS2 and SDPS2-SPMS heterodimers in vivo. The epitope-labeled SPDS and SPMS proteins copurified with protein complexes ranging in size from 650 to 750 kD. Our data demonstrate the existence of a metabolon involving at least the last two steps of polyamine biosynthesis in Arabidopsis.
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000166:cytology / Q000201:enzymology* / Q000235:genetics; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000378:metabolism; D001665:Binding Sites; D002478:Cells, Cultured; D000939:Epitopes / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D009154:Mutation; D010802:Phylogeny; D037121:Plant Lectins / Q000378:metabolism; D011073:Polyamines / Q000378:metabolism*; D011233:Precipitin Tests; D011485:Protein Binding; D025941:Protein Interaction Mapping; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism; D011700:Putrescine / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D013095:Spermidine / Q000096:biosynthesis; D011436:Spermidine Synthase / Q000235:genetics* / Q000378:metabolism*; D020798:Two-Hybrid System Techniques
D029681:Arabidopsis Proteins; D000939:Epitopes; D037121:Plant Lectins; D011073:Polyamines; D016271:Proto-Oncogene Proteins c-myc; C470419:SPDS1 protein, Arabidopsis; C470420:SPDS2 protein, Arabidopsis; D011436:Spermidine Synthase; D013095:Spermidine; D011700:Putrescine