Nucleic Acids Research, 1992, Vol. 20, No. 13 3463-3469
© 1992
MOLECULAR BIOLOGY |
Insertion (sufB) in the anticodon loop or base substitution (sufC) in the anticodon stem of tRNAPro2 from Salmonella typhimurium induces suppression of frameshift mutations
Department of Microbiology, University of Umeå S-901 87 Umeå 1Department of Botany, Lilla Frescati 5, University of Stockholm Sweden 2National Cancer Centre Research Institute Tsukiji 5-chome, Chyo-ku, Tokyo, Japan
* To whom correspondence should be addressed
Received February 12, 1992. Revised May 5, 1992. Accepted May 5, 1992.
The dominant +1 frameshift suppressors sufA6, sufB1 and sufB2, in Salmonella typhlmurium act at runs of C and affect tRNAPro1, tRNAPro2 and tRNAPro2, respectively. A recessive +1 frameshift suppressor, sufC, has a similar suppressor specificity (Riddle,D.L., and Roth.J.R., Mol. Biol. 66, 483 and 495, 1972). We show that sufC strains harbour two frameshift suppressors of which one, sufX201, is allelic to sufB. We cloned the sufB+ wild type allele and by recombination in vivo the mutations sufB1, sufB2 and sufX201. Determination of the DNA sequence revealed that the sufB1 and sufB2 mutations result in an extra G in the anticodon loop of the minor tRNAPro2. The sufX201 mutation results in a base substitution (G43 to A43) in the anticodon stem of this tRNA. Although the sufB1 and sufB2 mutations were earlier shown to be dominant, the sufB+ wild type allele on multi copy plasmid inhibited the chromosomal sufB1, sufB2 and sufX201 mediated frameshift suppression but not that mediated by the dominant sufA6 mutation. These results are discussed in view of the possible coding specificity of these mutated tRNAs. The DNA sequence showed a potential consensus promoter sequence upstream of the structural gene for tRNAPro2 and downstream a dyad symmetrical structure followed by a T cluster, a possible rho-independent termination signal. The Salmonella tRNAPro2 gene is identical to the Escherichia coli counterpart reported by Komlne.Y. et al. (J. Mol. Biol. 212, 579–598, 1990). While the 5' flanking sequence similarity between the two species is about 83%, the similarity of the 3' flanking sequence is only 42%. Still, the Salmonella tRNAPro2 gene has a rho-independent transcriptional termination signal similar to the one present in E.coli tRNAPro2 gene.
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