Biopolym. Cell. 1988; 4(1):3-7.
Structure and Function of Biopolymers
Thermodynamics of interaction of deacylated initiator tRNA and aminoacyl-tRNA with P- and A-sites of 30S subunits of Escherichia coli ribosomes programmed by AUG(U)n
- B. P. Konstantinov Institute of Nuclear Physics, Academy of Sciences of the USSR
Gatchina, Leningrad distr., USSR
Small ribosomal subunit of E. coli ribosome programmed by AUG(U)n is capable to bind two molecules of tRNA (tRNA,Met or fMet-tRNAfMet plus Phe-tRNAPhe). tRNAfMet is bound to P-site of SOS subunit of the complex [30S+AUG(U)n] (or to the site overlapped with P-site). Phe-tRNAphe is bound to A-site of 30S subunits of the complex [30S + 4-AUG(U)n+tRNAfMet] and its binding is inhibited by tetracycline. The association constants tRNAfMet with the complex [30S+AUG(U)n] and Phe-tRNAphe with the complex [30S+AUG(U)n+tRNAfMet] are measured at different temperatures and magnesium concentrations and thermodynamic parameters of these interactions are calculated. When fMet-tRNAfMet is bound instead of tRNAtMet, dipeptide (fMet-Phe) formation is observed after addition of SOS subunits. Thus, there are two tRNA binding sites on 30 S subunits which are realized as functional P- and A-sites of 70S ribosome not only in the poly(U)-dependent system but in the studied system with matrix carrying an initiator codon AUG.
 Adams JM, Capecchi MR. N-formylmethionyl-sRNA as the initiator of protein synthesis. Proc Natl Acad Sci U S A. 1966;55(1):147-55.
 Bretscher MS, Marcker KA. Polypeptidyl-sigma-ribonucleic acid and amino-acyl-sigma-ribonucleic acid binding sites on ribosomes. Nature. 1966;211(5047):380-4.
 Lucas-Lenard J, Lipmann F. Initiation of polyphenylalanine synthesis by N-acetylphenylalanyl-SRNA. Proc Natl Acad Sci U S A. 1967;57(4):1050-7.
 Shine J, Dalgarno L. Determinant of cistron specificity in bacterial ribosomes. Nature. 1975;254(5495):34-8.
 Stormo GD, Schneider TD, Gold LM. Characterization of translational initiation sites in E. coli. Nucleic Acids Res. 1982;10(9):2971-96.
 Katunin VI, Semenkov YP, Makhno VI, Kirillov SV. Comparative study of the interaction of polyuridylic acid with 30S subunits and 70S ribosomes of Escherichia coli. Nucleic Acids Res. 1980;8(2):403-21.
 Clark B. F. C. Structure of tRNA during protein biosynthesis. Ribosomes: structure, function and genetics . Eds G. Chambliss et al. Baltimore: Univ. Park press, 1980:413-443.
 Ofengand J. The topography of tRNA binding sites on the ribosomes. Eds G. Chambliss et al. Baltimore: Univ. Park press, 1980:497-529.
 Semenkov YuP, Makarov EM, Kirillov SV. Quantitative study of interaction of deacylated tRNA with the P, A and E sites of Escherichia coli ribosomes. Biopolym Cell. 1985; 1(4):183-93.
 Walker RT, RajBhandary UL. Studies on polynucleotides. CI. Escherichia coli tyrosine and formylmethionine transfer ribonucleic acids: effect of chemical modification of 4-thiouridine to uridine on their biological properties. J Biol Chem. 1972;247(15):4879-92.
 Aoyagi S, Inoue Y. Oligonucleotide studies. I. Isolation of dinucleotides and seven trinucleotides from ribonuclease T-1 digests of nucleic acids and their ultraviolet spectral characterizations. J Biol Chem. 1968;243(3):514-20.
 Szer W, Kurylo-Borowska Z. Effect of edeine on aminoacyl-tRNA binding to ribosomes and its relationship to ribosomal binding sites. Biochim Biophys Acta. 1970;224(2):477-86.
 Semenkov YuP, Makarov EM, Makhno VI, Kirillov SV. Kinetic aspects of tetracycline action on the acceptor (A) site of Escherichia coli ribosomes. FEBS Lett. 1982;144(1):125-9.
 Schmitt M, Neugebauer U, Bergmann C, Gassen HG, Riesner D. Binding of tRNA in different functional states to Escherichia coli ribosomes as measured by velocity sedimentation. Eur J Biochem. 1982;127(3):525-9.
 Kirillov SV. Codon-anticodon mechanism of interaction in ribosomes. Results of science and technology. Moscow, VINITI, 1983; 5-98 (Biol. Chemistry, Vol. 18).
 Ganoza MC, Sullivan P, Cunningham C, Hader P, Kofoid EC, Neilson T. Effect of bases contiguous to AUG on translation initiation. J Biol Chem. 1982;257(14):8228-32.