Translation error rate with protamine mRNA as a template

Range 0.0006-0.002 mismatched/matched aa
Organism Trout Oncorhynchus mykiss
Reference Mori N, Funatsu Y, Hiruta K, Goto S. Analysis of translational fidelity of ribosomes with protamine messenger RNA as a template. Biochemistry. 1985 Feb 26 24(5):1231-9. abstract & p.1237 left column 2nd paragraphPubMed ID4096904
Method Abstract: "A novel method was developed to estimate the translational fidelity of mammalian ribosomes in vitro with protamine mRNA of rainbow trout as template. Protamines are mixtures of basic proteins consisting of only seven types of amino acids (Arg, Ile, Val, Ser, Pro, Ala, and Gly), arginine (codon, AGR and CGN) being abundant. Taking advantage of the absence of lysine (codon, AAG) in the proteins, [investigators] determined the misincorporation of this amino acid into protamines in a cell-free translation system consisting of mouse liver ribosomes, protamine mRNA, [3H]lysine, [14C]arginine, and seven unlabeled amino acids: Ile, Val, Ser, Pro, Ala, Gly, and Met. After the reaction, translation products were analyzed by either sucrose gradient centrifugation or polyacrylamide gel electrophoresis." P.1237 left column 3rd paragraph: "[Researchers] developed an in vitro assay method for estimating the translational fidelity of eukaryotic ribosomes using protamine mRNA of rainbow trout."
Comments Abstract: "The frequency was found to be 0.0006-0.002. This method enabled [investigators] to determine the frequency of misrecognition of purine bases at the second position of arginine codons in mRNA." P.1237 left column 2nd paragraph: "[Investigators] calculated the lysine/arginine substitution frequency from the radioactivity in the slices containing protamine (Table IV). Three independent electrophoretic analyses were made, and calculations were made from results using different conditions for preparation of ribosomes and counting of radio-activity (see Experimental Procedures). The molar ratio of lysine/arginine was again found to be 0.00063-0.00195, values being of the same order of magnitude as the ratio observed on sucrose gradient analysis (Table II)." See BNID 103454
Entered by Uri M
ID 105718