The intermolecular complex of erythromycin and ribosome

Mao, James C.-H.; Putterman, Mary

doi:10.1016/0022-2836(69)90180-6

Cited by 94 publications

(35 citation statements)

References 31 publications

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“…From these studies, we infer that the N8-methyl group may weaken the participation of adenine in hydrogen-bond formation. Consistent with the importance of the role of the 6-amino group in erythromycin binding is the observation of Mao and Putterman (13) that treatment of 50S ribosomal subunits with formaldehyde greatly reduced the ability of these subunits to bind erythromycin. It is also of interest that the presence of dimethyladenylate residues in the 16S rRNA of E. coli was found by Fellner (10) to markedly alter the reactivity of oligonucleotides containing these groups in enzymatic and alkaline hydrolysis reactions, and he postulated that this effect might be mediated by (unspecified) alterations in configuration and/or charge properties of neighboring bases.…”

Section: Discussionsupporting

confidence: 68%

Altered Methylation of Ribosomal RNA in an Erythromycin-Resistant Strain of Staphylococcus aureus

Lai

Weisblum

1971

Proc. Natl. Acad. Sci. U.S.A.

252

114

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In certain strains of Staphylococcus aureus, a concentration of erythromycin between 10-8 and 10-7 M can induce resistance to concentrations of this drug as high as 10-4 M. In one such strain studied, S. aureus (1206), N6-dimethyladenine is not normally present in 23S rRNA; however, a compound presumptively identified (on the basis of paper chromatography in three different solvents) as N6-dimethyladenine appears in the 23S rRNA of growing cells that have been incubated in a medium containing 10-7 M erythromycin. It has been shown previously that the induction of the erythromycinresistant phenotype that occurs under these conditions requires 10-8-10-7 M erythromycin for maximal expression within 1 hr and that induction results in modified 50S ribosomal subunits, which are then unable to bind erythromycin or lincomycin. Methylated adenine is also found in the 16S rRNA from the strain of S. aureus studied; however, in contrast to the situation with 23S rRNA, the amount in 16S rRNA is not affected by erythromycin. These findings provide the first example of a correlation between the methylation of rRNA and altered ribosomal function.In certain strains of Staphylococcus aureus, erythromycin can function as an inducer of resistance to 3 of 8 classes of inhibitors known to act on the 50S ribosomal subunit; these include the macrolides, lincosamides, and streptogramin B families of antibiotics. Some of the properties of these inducible strains and of derived constitutive mutants have been described, together with references to the work of others (1, 2).50S ribosomal subunits obtained from induced-and constitutively-resistant cells show a decreased ability to bind erythromycin and lincomycin, and the possibility of enzymatic inactivation of these drugs was rigorously excluded. Previous studies suggested that erythromycin can induce a modification in the structure of the ribosome, and that synthesis of protein and RNA, but not of DNA, was required for this modification to be expressed. We therefore compared the structural components of sensitive and resistant ribosomes in order to localize the putative structural modification(s) induced by erythromycin. We found that a compound, tentatively identified as N6-dimethyladenine, was present in 23S rRNA obtained from induced-and constitutively-resistant cells, but not in 23S rRNA obtained from sensitive, uniniduced cells. MATERIALS AND METHODS Bacterial strains and mediumThe properties of S. aureus (strain 1206+), inducibly resistant to erythromycin, and of a derived constitutivelyresistant mutant have been described (1, 2). Preparation of ribosomes, subunits, and ribosomal RNA Cells were grown, harvested in the late-logarithmic phase, and stored as a frozen paste for 2 days at -50'C. The cell paste was suspended in standard buffer (10 maM Trise HCI (pH 7.4)-10 mM MgCl2-60 mM KCl-7 mM mercaptoethanol), mixed with two times its volume of glass beads (120-lim diameter), and disrupted with a Buhler mill. Nearly complete disruption was found after 45 min of treatment. The ...

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Section: Discussionsupporting

confidence: 68%

Altered Methylation of Ribosomal RNA in an Erythromycin-Resistant Strain of Staphylococcus aureus

Lai

Weisblum

1971

Proc. Natl. Acad. Sci. U.S.A.

252

114

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“…This phenomenon could be explained by considering some conformational change of the ribosomes by heat producing a lowered affinity for macrolides. According to Teraoka [22], some conformational structure of ribosomes from E. coli Q13 must be necessary for the expression of EM-binding, and Mao and Putterman [10] stated that 50S ribosomes of S. aureus 209P were found to bind EM but neither 42S nucleoprotein nor 50S protein bound the antibiotic. These facts also may support the above speculation of some altered structure of the heat-treated ribosomes, and it is suspected that viability might not directly affect the accumulation of macrolides in cells because of the experimental results of accumulation proceeding with UV-irradiated cells, although Mao and Putterman [9] described that only viable cells accumulated EM from the observation with heat-or toluene-treated cells of S. aureus.…”

Section: Discussionmentioning

confidence: 99%

Decrease in Accumulation of Macrolide Antibiotics as a Mechanism of Resistance in Staphylococcus aureus

Yamagishi

Nakajima

Inoue

et al. 1971

Japanese Journal of Microbiology

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A marked difference of levels of accumulation of macrolide antibiotics (Mac) was found between Mac-sensitive (E642-1 and 209P) and Mac-resistant (M5642, MS57, U9, and 606) strains of Staphylococcus aureus by means of qualitative and quantitative bioassay methods. Though the levels of accumulation of the antibiotics in cells of Mac-resistant strains were only one-tenth that of the Mac-sensitives, satisfactory evidence for a relationship between the accumulation of the antibiotics and the degree of sensitivity to those drugs was established in a physiological sense from a kinetic measurement of Ks and v in the accumulation reaction of the antibiotics in the cells. The experiments with tritiated erythromycin or josamycin also support these results. It is suspected, by estimation of the accumulation in heated or UV-irradiated cells, that levels of accumulation of the antibiotics in cells of S. aureus could be reflected by a binding affinity of their ribosomes for these antibiotics. There was no system for inactivating the antibiotics seen in the cells of the Mac-resistants, i.e., MS642, 606, and U9.

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“…Erythromycin and other macrolide antibiotics in particular have been widely studied for their effects on the functions of the ribosome during translation (12). These compounds bind strongly to the 50S ribosomal subunit of both gram-positive (12,35) and gram-negative (46) cells and interfere with the elongation of the nascent peptide chain (9,13,52).…”

mentioning

confidence: 99%

Ribosomal protein gene sequence changes in erythromycin-resistant mutants of Escherichia coli

1994

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The genes for ribosomal proteins IA and L22 from two erythromycin-resistant mutants of Escherichia coli have been isolated and sequenced. In the L4 mutant, an A-to-G transition in codon 63 predicted a Lys-to-Glu change in the protein. In the L22 strain, a 9-bp deletion removed codons 82 to 84, eliminating the sequence Met-Lys-Arg from the protein. Consistent with these DNA changes, in comparison with wild-type proteins, both mutant proteins had reduced first-dimension mobilities in two-dimensional polyacrylamide gels. Complementation of each mutation by a wild-type gene on a plasmid vector resulted in increased erythromycin sensitivity in the partial-diploid strains. The fraction of ribosomes containing the mutant form of the protein was increased by growth in the presence of erythromycin. Erythromycin binding was increased by the fraction of wild-type protein present in the ribosome population. The strain with the IA mutation was found to be cold sensitive for growth at 200C, and SOS-subunit assembly was impaired at this temperature. The mutated sequences are highly conserved in the corresponding proteins from a number of species. The results indicate the participation of these proteins in the interaction of erythromycin with the ribosome.The interaction of antimicrobial agents with the bacterial ribosome has been a significant area of research for many years because of its relevance to infectious diseases (20, 39). Erythromycin and other macrolide antibiotics in particular have been widely studied for their effects on the functions of the ribosome during translation (12). These compounds bind strongly to the 50S ribosomal subunit of both gram-positive (12, 35) and gram-negative (46) cells and interfere with the elongation of the nascent peptide chain (9, 13,52 (55). Affinity labeling studies with erythromycin derivatives have identified a strong interaction of the drug with protein L22 and weaker associations with proteins L2, IA, and L15 (2). Each of these proteins has been shown to be essential for-reconstitution of the peptidyltransferase activity of the 50S subunit (24). Recent studies have identified the location of several of these proteins at a common region in the 5OS-subunit structure (11).The involvement of rRNA in erythromycin activity has been indicated by a number of recent reports identifying 23S rRNA mutations leading to erythromycin resistance in Escherichia coli (15,18,50) (26,59).Resistance to erythromycin can also result from changes in ribosomal proteins in E. coli (1,(42)(43)(44)(45)58) and in Bacillus species (48,49,56). Whereas the RNA sequence changes have * Corresponding author. Phone: (615) 461-7040. been specifically identified, no sequence information about the alterations in ribosomal proteins leading to erythromycin resistance is presently available. Phenotypic effects of alterations in ribosomal proteins L4 and L22 in two resistant mutants of E. coli were described many years ago (1, 58). We have recently acquired these mutants and have determined the specific DNA sequence c...

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The intermolecular complex of erythromycin and ribosome

Cited by 94 publications

References 31 publications

Altered Methylation of Ribosomal RNA in an Erythromycin-Resistant Strain of Staphylococcus aureus

Altered Methylation of Ribosomal RNA in an Erythromycin-Resistant Strain of Staphylococcus aureus

Decrease in Accumulation of Macrolide Antibiotics as a Mechanism of Resistance in Staphylococcus aureus

Ribosomal protein gene sequence changes in erythromycin-resistant mutants of Escherichia coli

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