PMID: 6160468Jul 25, 1980Paper

Ribosome binding by tRNAs with fluorescent labeled 3' termini

Nucleic Acids Research
B D Wells, C R Cantor

Abstract

Yeast and E. coli tRNAPhe samples were oxidized and labeled at the 3' end with dansyl hydrazine or fluorescein thiosemicarbazide. These tRNAs can bind to poly(U)-programmed E. coli 70S tight couple ribosomes in 25 mM magnesium at 8 degrees C. Two binding sites with binding constants of about 1 X 10(9) M-1 (P) and 3 X 10(7) M-1 (A) were determined for the yeast tRNAPhe derivatives. With E. coli tRNAPhe the A site affinity is similar to yeast tRNAPhe but the P site affinity is 5-fold weaker. Singlet-singlet energy transfer showd that the distance from the 3' end of tRNAPhe in the P site to a fluorescein derivative of erythromycin is 23 A. This supports in vitro studies suggesting that erythromycin binds near the peptide moiety of peptidyl tRNA. A distance of 34 A between the 3' ends of 2 tRNAs bound simulatneously on the ribosome was also measured. This long distance may mean that the deacylated fluorescent tRNA binds to the A site in an orientation like that in the stringent response rather than in protein synthesis.

References

Jan 1, 1977·Nucleic Acids Research·B D Wells, C R Cantor
Oct 1, 1978·Nucleic Acids Research·V B Odinzov, S V Kirillov
Aug 25, 1979·Journal of Molecular Biology·R H FaircloughH G Zachau
Aug 25, 1979·Journal of Molecular Biology·R H Fairclough, C R Cantor
Jul 1, 1974·Proceedings of the National Academy of Sciences of the United States of America·C H Yang, D Söll
Jan 1, 1971·Annual Review of Biochemistry·J Lucas-Lenard
Dec 10, 1971·European Journal of Biochemistry·N De GrootY Lapidot

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Citations

Aug 27, 1990·Biochimica Et Biophysica Acta·B HardestyO W Odom
Apr 5, 1988·European Journal of Biochemistry·K FriedrichK G Steinhäuser
Jun 15, 1991·European Journal of Biochemistry·O W OdomB Hardesty
Mar 25, 1982·Journal of Molecular Biology·A E JohnsonC R Cantor

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