Translation in vitro of Tetrahymena pyriformis polyadenylated mRNA. Identification of tubulin amongst the translated products and demonstration of its heterogeneity.
The capacity of poly(A)-containing RNA of the protozoan ciliate Tetrahymena pyriformis to direct the synthesis of proteins in vitro has been tested using two cell-free systems: a wheat germ extract and a rabbit reticulocyte lysate. The results obtained with these two systems are compared and the identification of alpha and beta tubulins among the products of protein synthesis in vitro, after separation by one-dimensional and two-dimensional electrophoresis, is described. By isoelectric focusing in polyacrylamide gels, each species of tubulin is resolved into several bands, suggesting that the main subunits are more heterogeneous than has been generally described. Poly(A)-containing RNA has also been fractionated on a 70% formamide/sucrose gradient and it is shown that alpha and beta tubulins are coded by separate mRNAs.
D000818:Animals; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D010944:Plants / Q000378:metabolism; D011061:Poly A / Q000378:metabolism*; D011188:Potassium / Q000494:pharmacology; D014176:Protein Biosynthesis / Q000187:drug effects; D012333:RNA, Messenger / Q000378:metabolism*; D012156:Reticulocytes / Q000378:metabolism; D013095:Spermidine / Q000494:pharmacology*; D013769:Tetrahymena pyriformis / Q000378:metabolism*; D014908:Triticum / Q000378:metabolism; D014404:Tubulin / Q000096:biosynthesis*
D012333:RNA, Messenger; D014404:Tubulin; D011061:Poly A; D008274:Magnesium; D011188:Potassium; D013095:Spermidine