Enhanced differential synthesis of proteins in a mammalian cell-free system by addition of polyamines.
Addition of the polyamines spermidine, spermine, or putrescine to a fractionated mammalian cell-free protein-synthesis system programmed by a variety of mRNAs results in a 3- to 5-fold stimulation of amino acid incorporation over that found in the absence of added polyamine. The mRNAs used as template were adenovirus mRNA, globin 9s mRNA, and RNA from the bacteriophages R17, Qbeta, and MS2. The relative amounts of 10 adenovirus polypeptides synthesized in vitro are altered by the addition of polyamines to the translation system to reflect more closely the relative amounts of these polypeptides synthesized in vivo. This qualititive improvement in translation products on addition of polyamines allow the analysis of a number of products which are at best only marginally synthesized in the absence of added polyamines. The low level of synthesis due to endogenous mRNA is stimulated by spermidine and spermine but a lesser extent by putrescine.
D000818:Animals; D001201:Ascites; D002250:Carbon Radioisotopes; D002474:Cell-Free System; D002499:Centrifugation, Density Gradient; D002848:Chromatography, DEAE-Cellulose; D003090:Coliphages; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D007930:Leucine / Q000378:metabolism; D008274:Magnesium / Q000494:pharmacology; D008715:Methionine / Q000378:metabolism; D011073:Polyamines / Q000494:pharmacology; D014176:Protein Biosynthesis / Q000187:drug effects*; D011700:Putrescine / Q000494:pharmacology*; D012333:RNA, Messenger / Q000378:metabolism; D012335:RNA, Ribosomal / Q000378:metabolism; D011817:Rabbits; D012156:Reticulocytes; D013539:Simian virus 40; D012967:Sodium Dodecyl Sulfate; D013095:Spermidine / Q000494:pharmacology*; D013096:Spermine / Q000494:pharmacology*; D013462:Sulfur Radioisotopes; D013698:Templates, Genetic
D002250:Carbon Radioisotopes; D011073:Polyamines; D012333:RNA, Messenger; D012335:RNA, Ribosomal; D013462:Sulfur Radioisotopes; D013096:Spermine; D012967:Sodium Dodecyl Sulfate; D008715:Methionine; D007930:Leucine; D008274:Magnesium; D013095:Spermidine; D011700:Putrescine