Abstract
We report a novel mutation, C1066U in 16S rRNA which was selected for resistance to spectinomycin, an antibiotic which inhibits ribosomal translocation. The minimal inhibitory concentration (MIC) of spectinomycin determined for this mutant (15 micrograms/ml) is greater than with the wild-type plasmid (5 micrograms/ml) but lower than with the well known C1192U mutation (> 80 micrograms/ml). The C1066U mutation also increases the cells sensitivity to fusidic acid, another antibiotic which inhibits translation at the translocation stage, whereas C1192U is unchanged relative to the wild type. We discuss why the acquisition of resistance to one of these drugs is often associated with hypersensitivity to the other.
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- Bilgin N., Richter A. A., Ehrenberg M., Dahlberg A. E., Kurland C. G. Ribosomal RNA and protein mutants resistant to spectinomycin. EMBO J. 1990 Mar;9(3):735–739. doi: 10.1002/j.1460-2075.1990.tb08167.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bollen A., Davies J., Ozaki M., Mizushima S. Ribosomal Protein Conferring Sensitivity to the Antibiotic Spectinomycin in Escherichia coli. Science. 1969 Jul 4;165(3888):85–86. doi: 10.1126/science.165.3888.85. [DOI] [PubMed] [Google Scholar]
- Brake A. J., Fowler A. V., Zabin I., Kania J., Müller-Hill B. beta-Galactosidase chimeras: primary structure of a lac repressor-beta-galactosidase protein. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4824–4827. doi: 10.1073/pnas.75.10.4824. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brimacombe R. RNA-protein interactions in the Escherichia coli ribosome. Biochimie. 1991 Jul-Aug;73(7-8):927–936. doi: 10.1016/0300-9084(91)90134-m. [DOI] [PubMed] [Google Scholar]
- Brink M. F., Brink G., Verbeet M. P., de Boer H. A. Spectinomycin interacts specifically with the residues G1064 and C1192 in 16S rRNA, thereby potentially freezing this molecule into an inactive conformation. Nucleic Acids Res. 1994 Feb 11;22(3):325–331. doi: 10.1093/nar/22.3.325. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brosius J., Ullrich A., Raker M. A., Gray A., Dull T. J., Gutell R. R., Noller H. F. Construction and fine mapping of recombinant plasmids containing the rrnB ribosomal RNA operon of E. coli. Plasmid. 1981 Jul;6(1):112–118. doi: 10.1016/0147-619x(81)90058-5. [DOI] [PubMed] [Google Scholar]
- De Stasio E. A., Dahlberg A. E. Effects of mutagenesis of a conserved base-paired site near the decoding region of Escherichia coli 16 S ribosomal RNA. J Mol Biol. 1990 Mar 5;212(1):127–133. doi: 10.1016/0022-2836(90)90309-A. [DOI] [PubMed] [Google Scholar]
- Dontsova O., Dokudovskaya S., Kopylov A., Bogdanov A., Rinke-Appel J., Jünke N., Brimacombe R. Three widely separated positions in the 16S RNA lie in or close to the ribosomal decoding region; a site-directed cross-linking study with mRNA analogues. EMBO J. 1992 Aug;11(8):3105–3116. doi: 10.1002/j.1460-2075.1992.tb05383.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fromm H., Edelman M., Aviv D., Galun E. The molecular basis for rRNA-dependent spectinomycin resistance in Nicotiana chloroplasts. EMBO J. 1987 Nov;6(11):3233–3237. doi: 10.1002/j.1460-2075.1987.tb02640.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gutell R. R., Woese C. R. Higher order structural elements in ribosomal RNAs: pseudo-knots and the use of noncanonical pairs. Proc Natl Acad Sci U S A. 1990 Jan;87(2):663–667. doi: 10.1073/pnas.87.2.663. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Harris E. H., Burkhart B. D., Gillham N. W., Boynton J. E. Antibiotic resistance mutations in the chloroplast 16S and 23S rRNA genes of Chlamydomonas reinhardtii: correlation of genetic and physical maps of the chloroplast genome. Genetics. 1989 Oct;123(2):281–292. doi: 10.1093/genetics/123.2.281. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hou Y., Lin Y. P., Sharer J. D., March P. E. In vivo selection of conditional-lethal mutations in the gene encoding elongation factor G of Escherichia coli. J Bacteriol. 1994 Jan;176(1):123–129. doi: 10.1128/jb.176.1.123-129.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hui A. S., Eaton D. H., de Boer H. A. Mutagenesis at the mRNA decoding site in the 16S ribosomal RNA using the specialized ribosome system in Escherichia coli. EMBO J. 1988 Dec 20;7(13):4383–4388. doi: 10.1002/j.1460-2075.1988.tb03337.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Johanson U., Hughes D. Fusidic acid-resistant mutants define three regions in elongation factor G of Salmonella typhimurium. Gene. 1994 May 27;143(1):55–59. doi: 10.1016/0378-1119(94)90604-1. [DOI] [PubMed] [Google Scholar]
- Makosky P. C., Dahlberg A. E. Spectinomycin resistance at site 1192 in 16S ribosomal RNA of E. coli: an analysis of three mutants. Biochimie. 1987 Aug;69(8):885–889. doi: 10.1016/0300-9084(87)90216-1. [DOI] [PubMed] [Google Scholar]
- Mark L. G., Sigmund C. D., Morgan E. A. Spectinomycin resistance due to a mutation in an rRNA operon of Escherichia coli. J Bacteriol. 1983 Sep;155(3):989–994. doi: 10.1128/jb.155.3.989-994.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Miller J. H., Ganem D., Lu P., Schmitz A. Genetic studies of the lac repressor. I. Correlation of mutational sites with specific amino acid residues: construction of a colinear gene-protein map. J Mol Biol. 1977 Jan 15;109(2):275–298. doi: 10.1016/s0022-2836(77)80034-x. [DOI] [PubMed] [Google Scholar]
- Moazed D., Noller H. F. Interaction of antibiotics with functional sites in 16S ribosomal RNA. Nature. 1987 Jun 4;327(6121):389–394. doi: 10.1038/327389a0. [DOI] [PubMed] [Google Scholar]
- Neefs J. M., Van de Peer Y., De Rijk P., Goris A., De Wachter R. Compilation of small ribosomal subunit RNA sequences. Nucleic Acids Res. 1991 Apr 25;19 (Suppl):1987–2015. doi: 10.1093/nar/19.suppl.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Remme J., Margus T., Villems R., Nierhaus K. H. The third ribosomal tRNA-binding site, the E site, is occupied in native polysomes. Eur J Biochem. 1989 Aug 1;183(2):281–284. doi: 10.1111/j.1432-1033.1989.tb14925.x. [DOI] [PubMed] [Google Scholar]
- Richter Dahlfors A. A., Kurland C. G. Novel mutants of elongation factor G. J Mol Biol. 1990 Oct 20;215(4):549–557. doi: 10.1016/s0022-2836(05)80167-6. [DOI] [PubMed] [Google Scholar]
- Samaha R. R., O'Brien B., O'Brien T. W., Noller H. F. Independent in vitro assembly of a ribonucleoprotein particle containing the 3' domain of 16S rRNA. Proc Natl Acad Sci U S A. 1994 Aug 16;91(17):7884–7888. doi: 10.1073/pnas.91.17.7884. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sigmund C. D., Ettayebi M., Morgan E. A. Antibiotic resistance mutations in 16S and 23S ribosomal RNA genes of Escherichia coli. Nucleic Acids Res. 1984 Jun 11;12(11):4653–4663. doi: 10.1093/nar/12.11.4653. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Svab Z., Maliga P. Mutation proximal to the tRNA binding region of the Nicotiana plastid 16S rRNA confers resistance to spectinomycin. Mol Gen Genet. 1991 Aug;228(1-2):316–319. doi: 10.1007/BF00282483. [DOI] [PubMed] [Google Scholar]
- Triman K., Becker E., Dammel C., Katz J., Mori H., Douthwaite S., Yapijakis C., Yoast S., Noller H. F. Isolation of temperature-sensitive mutants of 16 S rRNA in Escherichia coli. J Mol Biol. 1989 Oct 20;209(4):645–653. doi: 10.1016/0022-2836(89)92000-7. [DOI] [PubMed] [Google Scholar]
- Willie G. R., Richman N., Godtfredsen W. P., Bodley J. W. Some characteristics of and structural requirements for the interaction of 24,25-dihydrofusidic acid with ribosome - elongation factor g Complexes. Biochemistry. 1975 Apr 22;14(8):1713–1718. doi: 10.1021/bi00679a025. [DOI] [PubMed] [Google Scholar]

