Polyamine requirement for microbial protein synthesis: structural specificity in cell-free systems of Escherichia coli.
Cell-free protein synthesis was performed with synthetic or natural mRNA in an E. coli system containing physiological concentrations of Ca2, Mg2 and either one or both of the two natural polyamines of E. coli, spermidine and putrescine, or corresponding homologues. Putrescine does not permit poly(U)-dependent poly(Phe) synthesis unless spermidine or nor-spermidine is added. Spermidine supports homopeptide synthesis sufficiently well, its effect being stimulated by putrescine or homologous diamines with increasing chain length from 4 to 7 carbon atoms. Diaminopropane completely inhibits the spermidine-activated system in a competitive manner. Translation of MS2 phage RNA is supported by putrescine, the rate and quality (read through to the termination signal) of translation is optimized by spermidine or triamine homologues. MS2 phage RNA translation is supported by spermidine, putrescine has no further stimulatory effect but diaminoheptane enhances the rate of translation. In this case, however, premature chain termination does occur. The results indicate that spermidine is necessary for optimal poly(U) and MS2 phage RNA translation, that the aminopropyl moiety is important for its function and that the remaining side chain can be extended from C4 to C8. Putrescine may cooperate with spermidine but its chain length is rather critical, it cannot substitute for spermidine. The results indicate that the polyamines facilitate mRNA/tRNA/ribosome interactions in a specific manner.
D000596:Amino Acids / Q000378:metabolism; D001426:Bacterial Proteins / Q000096:biosynthesis*; D002474:Cell-Free System; D003090:Coliphages / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism*; D007700:Kinetics; D010452:Peptide Biosynthesis; D011073:Polyamines / Q000378:metabolism*; D014176:Protein Biosynthesis; D011700:Putrescine / Q000378:metabolism; D012343:RNA, Transfer / Q000378:metabolism; D012367:RNA, Viral / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism; D013095:Spermidine / Q000378:metabolism
D000596:Amino Acids; D001426:Bacterial Proteins; D011073:Polyamines; D012367:RNA, Viral; D012343:RNA, Transfer; D013095:Spermidine; D011700:Putrescine