Translational Attenuation Mechanism of ErmB Induction by Erythromycin Is Dependent on Two Leader Peptides

Wang, Shasha; Jiang, Kai; Du, Xinyue; Lu, Yanli; Liao, Lijun; He, Zhiying; He, Weizhi

doi:10.3389/fmicb.2021.690744

Cited by 13 publications

(28 citation statements)

References 30 publications

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“…The plasmid pGEX-4T-3 (GE Healthcare) was used as the vector for the generation of the pGEX- ermBL - ermB’ - lacZ α reporter plasmid as described in a previous study [ 17 ]. pGEX- ermCL - ermC’ - lacZ α was constructed in this study.…”

Section: Methodsmentioning

confidence: 99%

“…The pGEX reporter plasmid was constructed as described in a previous study [ 17 ]. In short, the pGEX vector replaced some new multiple cloning site sequences (SmaI-KpnI-XbaI-AflII-XhoI-TthIIII) with sequences between BspMI and TthIIII.…”

Section: Methodsmentioning

confidence: 99%

“…The β-galactosidase assay method was used as described previously [ 17 ]. E. coli strains carrying the pGEX reporter plasmid were grown in LB until OD600 ≈ 0.2.…”

Section: Methodsmentioning

confidence: 99%

“…In addition, in our previous study, we also found that another leader peptide named ermBL 2 is present in the regulatory region and is critical for erythromycin (Ery)-mediated induction of gene expression (Fig. S 1 ) [ 17 ].…”

Section: Introductionmentioning

confidence: 99%

“…When the expression of ermB is inducible, ribosome stalling and mRNA stability are believed to control its expression [ 6 , 18 , 19 ]. In the Enterococcus faecalis strain DS16 transposon Tn917 (M11180) [ 16 ], ermB is preceded by a 258 nucleotide leader region, which contains two regulatory open reading frames, ermBL , which encodes a 27 amino acid-long leader peptide, and ermBL2 , which encodes a 16 amino acid-long leader peptide [ 6 , 17 , 19 , 20 ]. This regulatory region has been well studied in previous research [ 6 , 19 , 20 ].…”

Section: Introductionmentioning

confidence: 99%

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16-membered ring macrolides and erythromycin induce ermB expression by different mechanisms

Jiang

Qiu

et al. 2022

BMC Microbiol

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Background Ribosome stalling on ermBL at the tenth codon (Asp) and mRNA stabilization are believed to be mechanisms by which erythromycin (Ery) induces ermB expression. Expression of ermB is also induced by 16-membered ring macrolides (tylosin, josamycin and spiramycin), but the mechanism underlying this induction is unknown. Methods We introduced premature termination codons, alanine-scanning mutagenesis and amino acid mutations in ermBL and ermBL2. Results In this paper, we demonstrated that 16-membered ring macrolides can induce ermB expression but not ermC expression. The truncated mutants of the ermB-coding sequence indicate that the regulatory regions of ermB whose expression is induced by Ery and 16-membered ring macrolides are different. We proved that translation of the N-terminal region of ermBL is key for the induction of ermB expression by Ery, spiramycin (Spi) and tylosin (Tyl). We also demonstrated that ermBL2 is critical for the induction of ermB expression by erythromycin but not by 16-membered ring macrolides. Conclusions The translation of ermBL and the RNA sequence of the C-terminus of ermBL are critical for the induction of ermB expression by Spi and Tyl.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

“…The β-galactosidase assay method was used as described previously [ 17 ]. E. coli strains carrying the pGEX reporter plasmid were grown in LB until OD600 ≈ 0.2.…”

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

16-membered ring macrolides and erythromycin induce ermB expression by different mechanisms

Jiang

Qiu

et al. 2022

BMC Microbiol

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Regulation Transcriptional of Antibiotic Resistance Genes (ARGs) in Bacteria Isolated from WWTP

Díaz-Palafox,

Tamayo-Ordoñez,

Bello-López

et al. 2023

Curr Microbiol

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The incidence of antibiotics and transcriptional regulation of ARGs in isolated bacteria from wastewater needs to be explored. By HPLC, in samples of untreated wastewater, ampicillin (49.74 ± 5.70 µg/mL), chloramphenicol (0.60 ± 0.03 µg/mL), tylosin (72.95 ± 2.03 µg/mL), and oxytetracycline (0.22 ± 0.01 µg/mL) was determined. Through metagenomic analysis identified 58 bacterial species belonging to 9 phyla and at least 14 species have shown resistance to a variety of antibiotics. Twenty-two bacterial isolates were proved to be resistant to fifteen antibiotics of new generation and used in medical research to combat infectious diseases. Fourteen strains were shown to harbor plasmids in size ranges of 2–5 Kb, 6–10 Kb and plasmids with size greater than 10 Kb. By quantitative PCR it was possible to identify genes sul, qnr, cat1, aadA1, and sat-1 gene were shown to be present in gDNA samples from treated and untreated samples of wastewater and by relative expression analysis, differential expression of cat1, ermB, act, and tetA genes was demonstrated in strains that showed identity with Escherichia coli, Bacteroides fragilis, and Salmonella thyphi, and that were stressed with different concentrations of antibiotics. The presence of ARGs in untreated water samples, as well as in bacterial isolates, was indicative that in these habitats there are microorganisms that can resist β-lactams, aminoglycosides, tetracyclines, sulfonamides, and quinolones.

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Detection of Tylosin Resistance in Fusobacterium necrophorum subspecies necrophorum

Schwarz,

Mathieu,

Laverde Gomez

et al. 2024

ACS Agric. Sci. Technol.

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In-feed tylosin, a macrolide, is widely used to prevent liver abscessation in feedlot cattle by repressing growth of ruminal Fusobacterium necrophorum. Although tylosin has been used for almost five decades, no resistant F. necrophorum subsp. necrophorum strain has ever been isolated. Here, we report two strains (FN37 and FN38) previously isolated from abscessed livers containing several antibiotic resistance genes: cfr(C), tet(O), ant(6)-Ia, and erm(B), the latter of which confers resistance to macrolides via modification of the ribosome. To evaluate if erm(B) conferred a phenotypic advantage, four strains (deposited strain ATCC 25286, ruminal isolate FNC, and abscess isolates FN37 and FN38) were tested for their responses to tylosin. The two erm(B)-harboring strains showed resistance at concentrations commonly found within the ruminal compartment under current dosing guidelines, and in the case of FN38, up to 100 μg/mL tylosin was tolerated. Tylosin susceptibility varied depending on the growth phase (stationary vs logarithmic) and preconditioning (growth in medium containing tylosin at a concentration of 1 μg/mL) of the inoculum in all four strains, but the two harboring the erm(B) gene demonstrated robust resistance. This discovery along with whole genome sequencing and preliminary annotation indicates horizontal gene transfer and acquisition of resistance genes, highlighting the need to revisit antimicrobial strategies for the feedlot cattle industry.

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Translational Attenuation Mechanism of ErmB Induction by Erythromycin Is Dependent on Two Leader Peptides

Cited by 13 publications

References 30 publications

16-membered ring macrolides and erythromycin induce ermB expression by different mechanisms

16-membered ring macrolides and erythromycin induce ermB expression by different mechanisms

Regulation Transcriptional of Antibiotic Resistance Genes (ARGs) in Bacteria Isolated from WWTP

Detection of Tylosin Resistance in Fusobacterium necrophorum subspecies necrophorum

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