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Nucleic Acids Research, 1975, Vol. 2, No. 7 1177-1188
© 1975

Articles

The binding sites for tRNA on eukaryotic ribosomes

David P. Leader and Gordon C. Machray

Department of Biochemistry, University of Glasgow Glasgow G12 8QQ, UK

Received May 22, 1975. We have studied the non-enzymic binding of phe-tRNA to ribosomes from rat liver using deacylated tRNA to inhibit binding to the P-site and puromycin (5 x 10-3M) to inhibit binding to the A-site. We conclude that at a low concentration of magnesium ions (10mM) phe-tRKA is bound only at the A-site of 80S ribosomes, whereas at a high concentration of magnesium ions (40mM) phe-tRNA is also bound at the P-site. Studies with edeine indicate that, during non-enzymic binding of phe-tRNA, eukaryotic ribosomes (in contrast to prokaryotic ribosomes) have the A-site of the 60S subunit and the initiation site of the 40S subunit juxtaposed. This may account for the differences observed, in formation of diphenylalanyl-tRNA and phenylalanyl puromycin, between phe-tRNA bound non-enzymically to the P-sites of eukaryotic and prokaryotic ribosomes.


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