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Nucleic Acids Research, 1987, Vol. 15, No. 11 4669-4686
© 1987

Articles

The bases of the tRNA anticodon loop are independent by genetic criteria

Drew Smith, Linda Breeden*, Eleanor Farrell and Michael Yarus

Department of Molecular, Cellular and Developmental Biology, University of Colorado Boulder, CO 80309, USA

Received December 12, 1986. Revised April 19, 1987. Accepted April 19, 1987.

We employed two methods to study the translational role of interactions between anticodon loop nucleotides. Starting with a set of previously constructed weakly-suppressing anticodon loop mutants of Su7, we searched for second-site revertants that increase amber suppressor efficiency. Though hundreds of revertants were characterized, no second-site revertants were found in the anticodon loop. Second site reversion was detected in the D-stem, thereby demonstrating the efficacy of the search method. As a second method for detecting interactions, we used site-directed mutagenesis to construct multiple mutations in the anticodon loop. These multiple mutants are very weak suppressors and have translational activities that are equal to or lower than that predicted for the independent action of single mutations. We conclude that although the anticodon loop sequence of Su7 has an optimal structure for the translation of amber codons, we find no evidence that interactions between loop bases can enhance translational efficiency.

*Present address: MRC Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK


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