Identification and Characterization of a Novel Major Facilitator Superfamily Efflux Pump, SA09310, Mediating Tetracycline Resistance in Staphylococcus aureus

Li, Daiyu; Ge, Yan; Wang, Ning; Shi, Yunying; Guo, Gang; Zou, Quanming; Liu, Qiang

doi:10.1128/aac.01696-22

Cited by 6 publications

(4 citation statements)

References 41 publications

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“…3 ) 28 . When it came to inducing efflux activity and norA gene expression, CIP was more effective than DO at increasing norA gene expression and efflux activity 44 . According to the current study, TQ is more capable of reducing CIP-MIC than VP.…”

Section: Discussionmentioning

confidence: 92%

Thymoquinone’ potent impairment of multidrug-resistant Staphylococcus aureus NorA efflux pump activity

Ahmad,

Abdelgalil,

Khamis

et al. 2024

Sci Rep

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The drug efflux pump is a crucial mechanism implicated in resistance to multiple antimicrobials. Thymoquinone (TQ) has evidently demonstrated multiple activities, antibacterial being the most effective. Knowledge about TQ activity against multidrug-resistant Staphylococcus aureus is very scarce. Therefore, the present study was conducted to investigate TQ resistance modulation in ciprofloxacin (CIP) and doxycycline (DO) multidrug-resistant S. aureus. Forty-seven samples were collected from different sources, and S. aureus was isolated and identified. Then, S. aureus resistance profiles to antimicrobials, N. sativa essential oil, and TQ; the correlation between TQ-MIC readings and disc diffusion; cartwheel and ethidium bromide (EtBr) accumulation assays; and norA gene expression were all described within silico molecular docking for TQ interactions with norA efflux pump protein. TQ-MICs ranged from 5–320 µg/ml. TQ down-regulated norA gene expression, resulting in a drop in efflux pump activity of 77.5–90.6% in the examined strains, comparable to that observed with verapamil. Exposure of S. aureus strains to CIP and DO raises the initial basal efflux pumping expression to 34.2 and 22.9 times, respectively. This induced efflux pumping overexpression was substantially reduced by 97.7% when TQ was combined with CIP or DO. There was a significant reduction of MICs of CIP and DO MICs by 2–15 and 2–4 folds, respectively, after treatment with 0.5XMIC-TQ in resistance modulation assays. These results refer to TQ ligand inhibitory interactions with NorA protein in molecular docking. Interpretations of inhibition zone diameters (IZDs) of disc diffusion and TQ-MICs exhibit independence of MICs from IZDs, as indicated by invalid linear regression analysis. TQ significantly reduced efflux pumping S. aureus induced by CIP and DO, but further investigations are needed to improve TQ-pharmacokinetics to restore CIP and DO activity and suppress fluoroquinolone and doxycycline-resistant S. aureus selection in clinical and animal settings.

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Section: Discussionmentioning

confidence: 92%

Thymoquinone’ potent impairment of multidrug-resistant Staphylococcus aureus NorA efflux pump activity

Ahmad,

Abdelgalil,

Khamis

et al. 2024

Sci Rep

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“…Gene deletion mutants, including those with fruR (Δ sa03665 ), fruK (Δ sa03670 ), fruT (Δ sa03675 ), or fruKT (Δ sa03670_03675 ) knockout, were successfully constructed by means of homologous recombination, following the described methods [ 16 ]. The corresponding complementary strains of each knockout mutant were obtained by expressing the deleted gene under the control of a constitutive promoter using a replicative vector, namely pQLV1025 [ 15 ].…”

Section: Methodsmentioning

confidence: 99%

“…RNA isolation and RT-qPCR were conducted as previously described [ 16 ]. To analyze the expression levels of the fruRKT genes in SMM minimal medium supplemented with 0.5% glucose or fructose, bacterial cells in mid-exponential phase cultured in TSB medium were harvested by centrifugation and washed thrice with PBS.…”

Section: Methodsmentioning

confidence: 99%

Unveiling the fructose metabolism system in Staphylococcus aureus: insights into the regulatory role of FruR and the FruRKT operon in bacterial fitness

Ge,

Li,

Wang

et al. 2024

BMC Microbiol

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Background The utilization of fructose as a carbon source and energy provider plays a crucial role in bacterial metabolism. Additionally, fructose metabolism directly impacts the pathogenicity and virulence of certain pathogenic microorganisms. Results In this study, we report the discovery of a fructose phosphotransferase system (PTS) in S. aureus. This system comprises three genes, namely fruR, fruK, and fruT, which are co-located in an operon that is indispensable for fructose utilization in S. aureus. Our findings confirm that these three genes are transcribed from a single promoter located upstream of the fruRKT operon. The fruR gene encodes a DeoR-type transcriptional regulator, designated as FruR, which represses the expression of the fruRKT operon by direct binding to its promoter region. Significantly, our experimental data demonstrate that the fruRKT operon can be induced by fructose, suggesting a potential regulatory mechanism involving intracellular fructose-1-phosphate as a direct inducer. Furthermore, we conducted RNA-seq analysis to investigate the specificity of FruR regulation in S. aureus, revealing that the fruRKT operon is predominantly regulated by FruR. Conclusions In summary, this study has uncovered a fructose phosphotransferase system (PTS) in S. aureus, highlighting the essential role of the fruR, fruK, and fruT genes in fructose utilization. We confirmed their co-location within an operon and established FruR as a key regulator by binding to the operon’s promoter. Importantly, we demonstrated that fructose can induce this operon, possibly through intracellular fructose-1-phosphate. Our identification of this PTS system represents the initial characterization of a fructose metabolism system in S. aureus.

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“…TetK is typically housed on the small multicopy pT181 plasmid within the SCCmecIII of MRSA strains [87,88]. A novel MFS efflux pump, SA09310, has also recently been shown to mediate tetracycline (TET) resistance; a Δsa09310 mutant displays a 64-fold and 8-fold reduction in MIC against TET and doxycycline (DOX) [89]. The second is through the expression of ribosomal protection proteins (RPPs), TetO and TetM, which bind to the ribosome and prevent the access of TET to their target.…”

Section: Protein Synthesismentioning

confidence: 99%

Staph wars: the antibiotic pipeline strikes back

Douglas,

Laabei

2023

Microbiology

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Antibiotic chemotherapy is widely regarded as one of the most significant medical advancements in history. However, the continued misuse of antibiotics has contributed to the rapid rise of antimicrobial resistance (AMR) globally. Staphylococcus aureus , a major human pathogen, has become synonymous with multidrug resistance and is a leading antimicrobial-resistant pathogen causing significant morbidity and mortality worldwide. This review focuses on (1) the targets of current anti-staphylococcal antibiotics and the specific mechanisms that confirm resistance; (2) an in-depth analysis of recently licensed antibiotics approved for the treatment of S. aureus infections; and (3) an examination of the pre-clinical pipeline of anti-staphylococcal compounds. In addition, we examine the molecular mechanism of action of novel antimicrobials and derivatives of existing classes of antibiotics, collate data on the emergence of resistance to new compounds and provide an overview of key data from clinical trials evaluating anti-staphylococcal compounds. We present several successful cases in the development of alternative forms of existing antibiotics that have activity against multidrug-resistant S. aureus . Pre-clinical antimicrobials show promise, but more focus and funding are required to develop novel classes of compounds that can curtail the spread of and sustainably control antimicrobial-resistant S. aureus infections.

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Identification and Characterization of a Novel Major Facilitator Superfamily Efflux Pump, SA09310, Mediating Tetracycline Resistance in Staphylococcus aureus

Abstract: Drug efflux systems have recently been recognized as an important mechanism of multidrug resistance in bacteria. Here, we described the identification and characterization of a novel chromosomally encoded multidrug efflux pump (SA09310) in Staphylococcus aureus .

Cited by 6 publications

References 41 publications

Thymoquinone’ potent impairment of multidrug-resistant Staphylococcus aureus NorA efflux pump activity

Thymoquinone’ potent impairment of multidrug-resistant Staphylococcus aureus NorA efflux pump activity

Unveiling the fructose metabolism system in Staphylococcus aureus: insights into the regulatory role of FruR and the FruRKT operon in bacterial fitness

Staph wars: the antibiotic pipeline strikes back

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