Spermidine but not spermine is essential for hypusine biosynthesis and growth in Saccharomyces cerevisiae: spermine is converted to spermidine in vivo by the FMS1-amine oxidase.
In our earlier work we showed that either spermidine or spermine could support the growth of spe2Delta or spe3Delta polyamine-requiring mutants, but it was unclear whether the cells had a specific requirement for either of these amines. In the current work, we demonstrate that spermidine is specifically required for the growth of Saccharomyces cerevisiae. We were able to show this specificity by using a spe3Delta fms1Delta mutant that lacked both spermidine synthase and the FMS1-encoded amine oxidase that oxidizes spermine to spermidine. The polyamine requirement for the growth of this double mutant could only be satisfied by spermidine; i.e., spermine was not effective because it cannot be oxidized to spermidine in the absence of the FMS1 gene. We also showed that at least one of the reasons for the absolute requirement for spermidine for growth is the specificity of its function as a necessary substrate for the hypusine modification of eIF5A. Spermine itself cannot be used for the hypusine modification, unless it is oxidized to spermidine by the Fms1 amine oxidase. We have quantified the conversion of spermine in vivo and have shown that this conversion is markedly increased in a strain overexpressing the Fms1 protein. We have also shown this conversion in enzymatic studies by using the purified amine oxidase from yeast.
Chattopadhyay
Manas K
MK
Laboratory of Biochemistry and Genetics, Building 8, Room 223, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892, USA.
Tabor
Celia White
CW
Tabor
Herbert
H
D001483:Base Sequence; D004271:DNA, Fungal / Q000235:genetics; D005800:Genes, Fungal; D007700:Kinetics; D008239:Lysine / Q000031:analogs & derivatives* / Q000096:biosynthesis*; D009154:Mutation; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000235:genetics / Q000378:metabolism*; D010448:Peptide Initiation Factors / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D013095:Spermidine / Q000378:metabolism*; D013096:Spermine / Q000378:metabolism*; D013379:Substrate Specificity; D000097574:Eukaryotic Translation Initiation Factor 5A; D000097667:Polyamine Oxidase
D004271:DNA, Fungal; D008239:Lysine; D000587:Oxidoreductases Acting on CH-NH Group Donors; D010448:Peptide Initiation Factors; D016601:RNA-Binding Proteins; D029701:Saccharomyces cerevisiae Proteins; D013095:Spermidine; D013096:Spermine; D000097574:Eukaryotic Translation Initiation Factor 5A; D000097667:Polyamine Oxidase; C108504:FMS1 protein, S cerevisiae; C100028:hypusine