Bactericidal Antibiotics Induce Toxic Metabolic Perturbations that Lead to Cellular Damage

Belenky, Peter; Ye, Jonathan D.; Porter, Caroline; Cohen, Nadia; Lobritz, Michael A.; Ferrante, Thomas; Jain, Saloni R.; Korry, Benjamin J.; Schwarz, Eric G.; Walker, Graham C.; Collins, James J.

doi:10.1016/j.celrep.2015.09.059

Cited by 409 publications

(405 citation statements)

References 95 publications

Supporting

Mentioning

347

Contrasting

Unclassified

Order By: Relevance

“…It is the amount and target of Fenton oxidants that are relevant for antibiotic lethality, not the intracellular levels of endogenously produced O 2 − or H 2 O 2 . These and other results suggest that the incorporation of oxidized nucleotides into nascent DNA followed by incomplete base excision repair (BER) is an important molecular mechanism that contributes to the ROS-dependent component of antibiotic lethality (11,12,14,18). To date, no alternative interpretation has been suggested for this body of experimental observations.…”

Section: Significancementioning

confidence: 86%

“…It is particularly relevant that nucleoside triphosphates and nucleic acids are anionic ligands that complex with Fe +2 and promote the Fenton reaction to an extent comparable to EDTA and nitrilotriacetate (38,41). Furthermore, the proximity of the nucleic acid base of a dNTP or NTP to an Fe +2 complexed by its phosphates (42) favors its reaction with highly reactive Fenton oxidants (11,16). Since the diffusion distance for a hydroxyl radical is only one carbon bond length (43), nucleotides and nucleic acids must be at higher risk of damage from Fenton oxidants than many other biomolecules because of their ability to complex Fe +2 and promote local production of Fenton oxidants.…”

Section: Discussionmentioning

confidence: 99%

“…Third, we used an ELISA to show that MalE-LacZ induction leads to increased amounts of 8-oxo-guanine (8-oxo-7,8-dihydroguanine) in DNA (Fig. 2C), a well-established biomarker for oxidative stress (11) that is relevant to experiments described below. Fourth, we used two dyes based on different chemistries that have been widely used for ROS detection in previous studies (3, 12, 25): 3′-(p-hydroxyphenyl) fluorescein (HPF) and 5/6-chloromethyl-2′,7′-dichlorodihydrofluorescein diacetate (CM-H2DCFDA), whose fluorescence has been shown to correlate with O 2 − production determined by direct measurement (24).…”

Section: Male-lacz Induction Is Not Lethal In Anaerobic Conditions Bumentioning

confidence: 99%

“…This conclusion was challenged by critiques (8, 9) that focused particularly on an earlier paper that had carried out a systems-level analysis of the lethality of multiple classes of bactericidal antibiotics and proposed a model to account for the experimental observations (10). Substantial new evidence has subsequently been published that addresses concerns that were raised and strongly supports a contributing role for ROS in antibiotic lethality (11,12). Recent reviews have discussed how the apparently contradictory results can be explained in part by the specifics of the experimental setup and the technical details of the assays used (1, 6, 7).…”

mentioning

confidence: 99%

See 3 more Smart Citations

Lethality of MalE-LacZ hybrid protein shares mechanistic attributes with oxidative component of antibiotic lethality

Takahashi

Gruber

Yang

et al. 2017

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

View full text Add to dashboard Cite

Downstream metabolic events can contribute to the lethality of drugs or agents that interact with a primary cellular target. In bacteria, the production of reactive oxygen species (ROS) has been associated with the lethal effects of a variety of stresses including bactericidal antibiotics, but the relative contribution of this oxidative component to cell death depends on a variety of factors. Experimental evidence has suggested that unresolvable DNA problems caused by incorporation of oxidized nucleotides into nascent DNA followed by incomplete base excision repair contribute to the ROSdependent component of antibiotic lethality. Expression of the chimeric periplasmic-cytoplasmic MalE-LacZ 72-47 protein is an historically important lethal stress originally identified during seminal genetic experiments that defined the SecY-dependent protein translocation system. Multiple, independent lines of evidence presented here indicate that the predominant mechanism for MalE-LacZ lethality shares attributes with the ROS-dependent component of antibiotic lethality. MalE-LacZ lethality requires molecular oxygen, and its expression induces ROS production. The increased susceptibility of mutants sensitive to oxidative stress to MalE-LacZ lethality indicates that ROS contribute causally to cell death rather than simply being produced by dying cells. Observations that support the proposed mechanism of cell death include MalE-LacZ expression being bacteriostatic rather than bactericidal in cells that overexpress MutT, a nucleotide sanitizer that hydrolyzes 8-oxo-dGTP to the monophosphate, or that lack MutM and MutY, DNA glycosylases that process base pairs involving 8-oxo-dGTP. Our studies suggest stress-induced physiological changes that favor this mode of ROS-dependent death.A gents or conditions that inhibit important biological processes can kill cells. However, physiological processes metabolically downstream of the initial inhibition can also contribute to cell death (1). For example, β-lactam antibiotics not only inhibit penicillin-binding proteins leading to lysis; they also induce a futile cycle of cell wall synthesis and degradation that contributes to their killing (2). In another example, lethal attacks on Escherichia coli mediated by the type VI secretion system P1vir phage and the antimicrobial peptide polymyxin B elicit the production of reactive oxygen species (ROS) that contribute to cell death (3). In bacteria, ROS production has been associated with the lethal effects of diverse stresses (4). In most cases, the detailed mechanisms responsible for ROS production are poorly understood; however, a variety of futile metabolic cycles can elicit H 2 O 2 production, illustrating the breadth of possible metabolic perturbations that could potentially induce oxidative stress (5).Despite widespread evidence that endogenous ROS produced as a consequence of metabolic stress can be lethal to bacteria and eukaryotes, the application of this concept to antibiotic lethality has been complicated. Evidence from multiple investiga...

show abstract

Section: Significancementioning

confidence: 86%

Section: Discussionmentioning

confidence: 99%

Section: Male-lacz Induction Is Not Lethal In Anaerobic Conditions Bumentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

Lethality of MalE-LacZ hybrid protein shares mechanistic attributes with oxidative component of antibiotic lethality

Takahashi

Gruber

Yang

et al. 2017

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

View full text Add to dashboard Cite

show abstract

“…A recent study showed that, to survive sub-lethal concentrations of antibiotics, E. coli cells up-regulate their DNA/RNA repair (Belenky et al, 2015). This is thought to drain the nucleotide pools as they are immediately used for DNA/ RNA repair.…”

Section: Discussionmentioning

confidence: 99%

The anti-cancerous drug doxorubicin decreases the c-di-GMP content in Pseudomonas aeruginosa but promotes biofilm formation

et al. 2016

View full text Add to dashboard Cite

Current antibiotic treatments are insufficient in eradicating bacterial biofilms, which represent the primary cause of chronic bacterial infections. Thus, there is an urgent need for new strategies to eradicate biofilm infections. The second messenger c-di-GMP is a positive regulator of biofilm formation in many clinically relevant bacteria. It is hypothesized that drugs lowering the intracellular level of c-di-GMP will force biofilm bacteria into a more treatable planktonic lifestyle. To identify compounds capable of lowering c-di-GMP levels in Pseudomonas aeruginosa, we screened 5000 compounds for their potential c-di-GMP-lowering effect using a recently developed c-di-GMP biosensor strain. Our screen identified the anti-cancerous drug doxorubicin as a potent c-di-GMP inhibitor. In addition, the drug decreased the transcription of many biofilm-related genes. However, despite its effect on the c-di-GMP content in P. aeruginosa, doxorubicin was unable to inhibit biofilm formation or disperse established biofilms. On the contrary, the drug was found to promote P. aeruginosa biofilm formation, possibly through release of extracellular DNA from a subpopulation of killed bacteria. Our findings emphasize that lowering of the c-di-GMP content in bacteria might not be sufficient to mediate biofilm inhibition or dispersal.

show abstract

Medicinal Chemistry of β‐Lactam Antibiotics

Testero

Llarrull

Fisher

et al. 2021

Burger's Medicinal Chemistry and Drug Discovery

View full text Add to dashboard Cite

The β‐lactam class of antibacterials is a cornerstone of human health. For nearly eight decades, their unparalleled clinical efficacy and clinical safety have made the β‐lactam class preeminent in the treatment of bacterial infection. The relatively brief period in human history during which the β‐lactams have exerted this benefit is a period characterized by continuous medicinal chemistry innovation, seen visibly in the progression from the penicillins to the complex ensemble of β‐lactams (now including also cephalosporins, monobactams, and carbapenems) used in the clinic. The key force behind this innovation is the progressive evolution by bacteria of resistance mechanisms. Today, highly resistant bacteria challenge the way medicinal chemists contemplate the creative alteration of β‐lactam structures, the way the pharmaceutical industry develops β‐lactams (and other antibacterial) structures, and the way the medical community uses antibacterials. This article gives a concise summary of the history of the β‐lactams. Its emphasis is recent structural innovation with respect to the β‐lactams, and with respect to structurally related classes that act to preserve the clinical activity of the β‐lactams through inhibition of bacterial β‐lactam‐hydrolyzing, and thus β‐lactam‐deactivating, enzymes. We integrate these chemistry advances with new biological discoveries with respect to the bactericidal mechanism of the β‐lactams and with respect to bacterial resistance mechanisms. The combination of these perspectives is a foundational perspective to guide the medicinal chemistry future of the β‐lactams.

show abstract

Bactericidal Antibiotics Induce Toxic Metabolic Perturbations that Lead to Cellular Damage

Cited by 409 publications

References 95 publications

Lethality of MalE-LacZ hybrid protein shares mechanistic attributes with oxidative component of antibiotic lethality

Lethality of MalE-LacZ hybrid protein shares mechanistic attributes with oxidative component of antibiotic lethality

The anti-cancerous drug doxorubicin decreases the c-di-GMP content in Pseudomonas aeruginosa but promotes biofilm formation

Medicinal Chemistry of β‐Lactam Antibiotics

Contact Info

Product

Resources

About