Translation of poly(riboadenylic acid)-enriched messenger RNAs from the yeast, Saccharomyces cerevisiae, in heterologous cell-free systems.
Poly(riboadenylic acid) [poly(A)] enriched messenger RNAs from yeast have been used to direct the synthesis of yeast polypeptides in mouse Krebs II ascites and wheat embryo extracts. Both cell-free systems, synthesize polypeptides over a molecular weight range of 10,000-100,000. Autoradiograms of sodium dodecyl sulfate-polyacrylamide slab gels used to fractionate [35S]methionine-labeled in vitro products reveal that about 25 major bands (each of them possibly representing multiple polypeptides) are produced by each cell-free system. Each of these coelectrophoreses with a major polypeptide labeled in vivo or in a yeast lysate. These results suggest that cell-free translational machinery from eukaryotes is not able to discriminate in an all or none fashion against messenger RNAs which are available to it. While yeast poly(A)-enriched messenger RNA directs the synthesis polypeptides over approximately the same molecular weight range in both cell-free systems, the wheat germ system directs the incorporation of 45 times the amount of [3H]serine into Cl3CCOOH-precipitable polypeptides. This is in contrast to the 2.5-fold more efficient translation of hemoglobin mRNA in the wheat embryo extract. Thus, the extract from mammalian cells is able to translate mRNA from a lower plant with a much lower efficiency than it translates hemoglobin mRNA, and at a lower efficiency than is observed using a cell-free system from wheat embryos. This indicates that the wheat embryo system is the one of choice for translation of yeast messenger RNA.
D000227:Adenine Nucleotides; D000818:Animals; D001483:Base Sequence; D002287:Carcinoma, Krebs 2 / Q000378:metabolism; D002474:Cell-Free System; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D051379:Mice; D008970:Molecular Weight; D010452:Peptide Biosynthesis; D010944:Plants / Q000378:metabolism; D011188:Potassium / Q000494:pharmacology; D014176:Protein Biosynthesis / Q000187:drug effects; D012333:RNA, Messenger / Q000378:metabolism*; D011817:Rabbits; D012156:Reticulocytes / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D014908:Triticum / Q000378:metabolism
D000227:Adenine Nucleotides; D012333:RNA, Messenger; D008274:Magnesium; D011188:Potassium