Eukaryotic initiation factor 4D, the hypusine-containing protein, is conserved among eukaryotes.
When mammalian cells are grown in medium containing [3H]spermidine, a single major tritiated protein identical to eukaryotic initiation factor 4D becomes labeled. This protein contains 1 residue/molecule of tritiated hypusine (N epsilon-(4-amino-2-hydroxybutyl)lysine), a rare amino acid which has been found in no other protein. In order to investigate the conservation of this protein, we examined two nonmammalian eukaryotes, the yeast Saccharomyces cerevisiae and the insect Drosophila melanogaster, and the eubacterial prokaryote Escherichia coli for the presence of the hypusine-containing protein. When the eukaryotic cells were grown in the presence of [3H]spermidine, electrophoretic analysis revealed a single labeled protein. In each case, the apparent molecular weight was near 18,000 and the relative pI was approximately 5.2, similar to the hypusine-containing protein of mammals. Amino acid analysis confirmed the presence of tritiated hypusine in each case, and silver staining of two-dimensional polyacrylamide gels demonstrated that, in yeast and fruit flies as in mammals, the protein is relatively abundant. In the eubacterium E. coli, one tritiated protein was predominant, but its molecular weight was 24,000 and we found no evidence that it contained tritiated hypusine. We found no evidence for the existence of the hypusine-containing protein in the archaebacterium Methanococcus voltae. These data suggest that the hypusine-containing protein is conserved among eukaryotes.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002477:Cells / Q000032:analysis*; D004331:Drosophila melanogaster / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D005057:Eukaryotic Cells / Q000032:analysis* / Q000378:metabolism; D007526:Isoelectric Point; D008239:Lysine / Q000031:analogs & derivatives* / Q000032:analysis; D008970:Molecular Weight; D010448:Peptide Initiation Factors / Q000032:analysis*; D010449:Peptide Mapping; D016601:RNA-Binding Proteins; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D013095:Spermidine / Q000378:metabolism; D000097574:Eukaryotic Translation Initiation Factor 5A
D000596:Amino Acids; D008239:Lysine; D010448:Peptide Initiation Factors; D016601:RNA-Binding Proteins; D013095:Spermidine; D000097574:Eukaryotic Translation Initiation Factor 5A; C100028:hypusine