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Nucleic Acids Research, 1995, Vol. 23, No. 5 779-784
© 1995

RNA

Genetic selection for active E.coli amber tRNAAsn exclusively led to glutamine inserting suppressors

Franck Martin, Gilbert Eriani, Joseph Reinbolt, Guy Dirheimer and Jean Gangloff*

Unité Propre de Recherche 9002 du Centre National de la Recherche Scientifique, Institut de Biologie Moléculaire et Cellulaire 15 rue René Descartes, 67084 Strasbourg, France

*To whom correspondence should be addressed

Received November 16, 1994. Revised January 27, 1995. Accepted January 27, 1995.

Suppressor tRNAs are useful tools for determining identity elements which define recognition of tRNAs in vivo by their cognate aminoacyl-tRNA synthetases. This study was aimed at the isolation of active amber tRNAAsn. Nineteen mutated tRNAAsnCUA having amber suppressor activity were selected by an in vivo genetic screen, and all exclusively inserted glutamine. From analysis of the different mutations it Is concluded that glutamine accepting activity was obtained upon reducing the interaction strength between the first base pair of thetRNAAsaCUA by direct or Indirect effects. Failure to Isolate tRNAAsaCUA suppressors charged with asparagine as well as other evolutionary related amino acids is discussed.


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