Comparison of fMet-tRNAf and Met-tRNAf from Escherichia coli and rabbit liver in initiation of hemoglobin synthesis.
A comparison has been made of the ability of the formylated and unformylated initiator tRNAs of Escherichia coli and rabbit liver to participate in a number of model reactions of protein synthesis. These reactions include: (a) formation of a ternary complex composed of the initiator tRNA, GTP, and initiation factor MP; (b) ApUpG-directed binding of the initiator tRNA to 40 S subunits with initiation factor Ml; (c) formation of the artificial dipeptide, methionylpuromycin; (d) formation of the natural initial globin dipeptide, methionylvaline; and (e) synthesis of sheep alpha and betaB-globin chains on reticulocyte polysomes from a type BB sheep. The results of these studies indicate that although the prokaryotic initiator tRNA species function efficiently in the partial reactions which involve only binding, the methionine donated by the prokaryotic tRNA is not incorporated efficiently into peptide linkage. This suggests that the initial high level of binding of the E. coli initiator tRNAs may be nonspecific, and that the structure of the tRNA itself is important for specific recognition by eukaryotic initiation factors. The effect of formylation on the effectiveness of the initiator tRNA is not clear; it reduces activity in ternary complex formation, does not affect ApUpG-directed binding to 40 S subunits, and increases the rate or extent of incorporation of methionine, or both, into methionylpuromycin and globin chains.
D000818:Animals; D004926:Escherichia coli / Q000378:metabolism*; D005561:Formates; D005914:Globins / Q000096:biosynthesis; D006454:Hemoglobins / Q000096:biosynthesis*; D007700:Kinetics; D008099:Liver / Q000378:metabolism*; D008715:Methionine; D010442:Peptide Chain Initiation, Translational; D011485:Protein Binding; D011691:Puromycin; D012329:RNA, Bacterial / Q000378:metabolism*; D012343:RNA, Transfer / Q000378:metabolism*; D011817:Rabbits; D012156:Reticulocytes / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism; D014633:Valine
D005561:Formates; D006454:Hemoglobins; D012329:RNA, Bacterial; D011691:Puromycin; D005914:Globins; D012343:RNA, Transfer; D008715:Methionine; D014633:Valine