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Nucleic Acids Research, 1977, Vol. 4, No. 8 2799-2809
© 1977

Articles

Search for revertants of the glutamine mischarging mutants of Escherichia coli su+3 tyrosine suppressor tRNA that are able to insert tyrosine at the site of amber mutation

Julio E. Celis*, Mike Squire+, Keld Kaltoft* and Eva Riisom*

*Division of Biostructural Chemistry, Chemistry Institute, Aarhus University Aarhus, Denmark +MRC Laboratory of Molecular Biology Cambridge, UK

Received May 19, 1977. We have isolated a bacterial amber mutation (nadam) that is suppressed by the tyrosine inserting suppressor su+3 but not by the glutamine (su+2, su+3 Al, su+3 G82 and su+3 AlG82), serine (su+1) and leucine (su+6) inserting suppressors. The su+7 suppressor which inserts glutamine and tryptophan1 also suppresses this mutation indicating that tryptophan, in addition to tyrosine, is accepted at the site of amber mutation. We have used this amber mutation to search for revertants of the su+3 glutamine mischarging mutants su+3 Al, su+3 G82 and su+3 AlG82 that are able to insert tyrosine at the site of amber mutation. Two types of revertants were found in the case of su+3 Al. One type corresponding to the true revertants Al -> G, and the other to the second site revertants C81 -> U (AlU81). The AlU81 revertant has been shown to insert both glutamine and tyrosine at the site of amber mutation. Only true revertants (G82 -> A) were obtained when su+3 G82 was analyzed. No revertants were obtained in the case of the su+3 AlG82. These results are discussed in relation to aminoacyl-tRNA recognition.


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