Biopolym. Cell. 1987; 3(3):157-160.
Short Communications
The number of tRNA-binding sites on 80S ribosomes and their subunits
1Rodnina M. V., 2Semenkov Yu. P.
  1. Institute of Molecular Biology and Genetics, Academy of Sciences of the Ukrainian SSR
    Kiev, USSR
  2. B. P. Konstantinov Institute of Nuclear Physics, Academy of Sciences of the USSR
    Gatchina, Leningrad distr., USSR

Abstract

The ability of the rabbit liver ribosomes and their subunits to form complexes with different types of tRNAPhe has been studied by the nitrocellulose membrane filtration technique. 80S ribosomes are shown to have two binding sites for aminoacyl- or peptidyl-tRNA, but three binding sites for non-acylated tRNA. The third (E) site of 80S ribosome is formed by its 60S subunit. The number of tRNA binding sites on 80S ribosomes or 40S subunits is constant at different Mg2+ concentrations (5–20 mM). The analysis of I/v2 vs. 1/C plot or Scatchard plot indicates thai binding of AcPhe-tRNAPhe to ribosomes is of a cooperative process. 60S subunit is shown to have one codon-independent binding site specific for non-acylated tRNA.

References

[1] Krayevsky AA, Kukhanova MK. The peptidyltransferase center of ribosomes. Prog Nucleic Acid Res Mol Biol. 1979;23:1-51.
[2] Parfenov DV, Saminski? EM. Deacylated tRNA binds with 50S subunits of Escherichia coli ribosomes at a special site not corresponding to the P'-site. Mol Biol (Mosk). 1985;19(5):1378-85.
[3] Kirillov SV, Semenkov YuP. Extension of Watson's model for the elongation cycle of protein biosynthesis. J Biomol Struct Dyn. 1986;4(2):263-9.
[4] Kirillov SV Mechanism of codon-anticodon interaction on ribosomes. VINITI, (Results of Science and Technology. Ser. B. Volume 18). 1983:5-98.
[5] Kirillov SV, Semenkov IuP. Interaction of tRNA with ribosomes. Mol Biol (Mosk). 1984;18(5):1249-63.
[6] Wettstein FO, Noll H. Binding of transfer ribonucleic acid to ribosomes engaged in protein synthesis: number and properties of ribosomal binding sites. J Mol Biol. 1965;11:35-53.
[7] Semenkov YuP, Kirillov SV, Stahl J. 40 S subunits from rat liver ribosomes contain two codon-dependent sites for transfer RNA. FEBS Lett. 1985;193(1):105-8.
[8] Falvey AK, Staehelin T. Structure and function of mammalian ribosomes. I. Isolation and characterization of active liver ribosomal subunits. J Mol Biol. 1970;53(1):1-19.
[9] Semenkov Yu. P., Makarov E. M., Kirillov S. V. Quantitative study of interaction of deacylated tRNA with the P, A and E sites of Escherichia coli ribosomes. Biopolym Cell. 1985; 1(4):183-193.
[10] Nirenberg M, Leder P. RNA Codewords and protein synthesis. The effect of trinucleotides upon the binding of sRNA to ribosomes. Science. 1964;145(3639):1399-407.
[11] Rheinberger HJ, Sternbach H, Nierhaus KH. Three tRNA binding sites on Escherichia coli ribosomes. Proc Natl Acad Sci U S A. 1981;78(9):5310-4.
[12] Kirillov SV, Semenkov YuP. Non-exclusion principle of Ac-Phe-tRNAPhe interaction with the donor and acceptor sites of Escherichia coli ribosomes. FEBS Lett. 1982;148(2):235-8.
[13] Peters M, Yarus M. Transfer RNA selection at the ribosomal A and P sites. J Mol Biol. 1979;134(3):471-91.
[14] Buisson M, Reboud AM, Dubost S, Reboud JP. Non-acylated tRNA binding on rat liver 60S subunits. Biochem Biophys Res Commun. 1979;90(2):634-40.