Antibiotic resistance mutations in 16S and 23S ribosomal RNA genes ofEscherichia coli

Abstract

Recombinant DNA and classic genetic procedures were used to map a spectinomycin resistance mutation to a 121 base pair region of a 16S RNA gene and a macrolide-lincosamide-streptogramin type B resistance mutation to a 32 base pair region of a 23S RNA gene. DNA sequence analysis of these regions revealed that spectinomycin resistance results from a C/G to T/A transition at position 1192 of a 16S RNA gene. Resistance to macrolide, lincosamide and streptogramin type B antibiotics results from an A/T to T/A transversion at position 2058 of a 23S RNA gene. The alteration in 16S RNA is in a sequence that can participate in alternate base pairing arrangements that have been proposed to be involved in the translocation process. The alteration in 23S RNA identifies sequences important to peptidyl transfer.

Differences in 23S ribosomal RNA mutations between wild-type and mutant macrolide-resistant Chlamydia trachomatis isolates

“…This form of resistance was first identified in yeast, where the mitochondrial operon was mutated at position A2058 of the large-subunit rRNA (8). Subsequently, similar phenotypes were obtained in Escherichia (E.) coli by the expression of mutant rRNA alleles from multiple-copy plasmids (9). Several years later, further reports of rRNA mutations that conferred macrolide resistance to clinical pathogens began to appear in the literature (9)(10)(11).…”

“…Subsequently, similar phenotypes were obtained in Escherichia (E.) coli by the expression of mutant rRNA alleles from multiple-copy plasmids (9). Several years later, further reports of rRNA mutations that conferred macrolide resistance to clinical pathogens began to appear in the literature (9)(10)(11). Mutations at positions 2057, 2058, 2059 and 2611 (E. coli numbering) in the peptidyl transferase region of 23S rRNA are considered to be important in the development of drug resistance against macrolides (12).…”

Structure and function of ribosomal RNA

“…The involvement of the 3'-terminus of the 16S rRNA in mRNA binding (Shine & Dalgarno, 1974;Steitz & Jakes, 1975) is now well-established, and resistance to several antibiotics has been traced to point mutations in 16S or 23S RNA (e.g. Garrett, 1983;Sigmund et al, 1984). The mutations in 23S RNA are implicated in peptidyl transfer, and as already mentioned above the same region of the 23S RNA structure has been identified in a cross-link to a peptidyl-tRNA affinity analogue (Barta et al, 1984).…”

Mutagenesis at the mRNA decoding site in the 16S ribosomal RNA using the specialized ribosome system in Escherichia coli.

“…Mutational analyses have so far not contributed to the assembly and verification of this model because mutations in the 16S rRNA structural gene are relatively rare and limited to mutations conferring antibiotic resistance (Sigmund et al, 1984;Noller, 1986, 1987;and reviewed in Noller, 1984). The rarity of 16S rRNA mutations is caused by the fact that there are seven, essentially identical, ribosomal RNA operons per haploid genome; hence recessive mutations in one of the operons are masked by the function of the remaining operons.…”

“…The rarity of 16S rRNA mutations is caused by the fact that there are seven, essentially identical, ribosomal RNA operons per haploid genome; hence recessive mutations in one of the operons are masked by the function of the remaining operons. Also, no selection scheme for obtaining mutants that map in the structural ribosomal RNA gene is available other than selection schemes for antibiotic resistance (Noller, 1984;Sigmund et al, 1984;Ettayebi et al, 1985).…”