Cardiolipin Prevents Membrane Translocation and Permeabilization by Daptomycin

Zhang, TianHua; Muraih, Jawad K.; Tishbi, Nasim; Herskowitz, Jennifer; Victor, Rachel L.; Silverman, Jared; Uwumarenogie, Stephanie; Taylor, Scott D.; Palmer, Michaël; Mintzer, Evan

doi:10.1074/jbc.m114.554444

Cited by 141 publications

(171 citation statements)

References 27 publications

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“…SMRT sequencing has been used to characterize the methylomes of bacteria (47)(48)(49)(50)(51)(52)(53)(54). During SMRT sequencing, DNA polymerase kinetics are monitored in real time; modified bases in the sequencing template lead to altered polymerase kinetics relative to those obtained with an unmodified template (55,56).…”

Section: Resultsmentioning

confidence: 99%

Genome Modification in Enterococcus faecalis OG1RF Assessed by Bisulfite Sequencing and Single-Molecule Real-Time Sequencing

et al. 2015

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Enterococcus faecalis is a Gram-positive bacterium that natively colonizes the human gastrointestinal tract and opportunistically causes life-threatening infections. Multidrug-resistant (MDR) E. faecalis strains have emerged, reducing treatment options for these infections. MDR E. faecalis strains have large genomes containing mobile genetic elements (MGEs) that harbor genes for antibiotic resistance and virulence determinants. Bacteria commonly possess genome defense mechanisms to block MGE acquisition, and we hypothesize that these mechanisms have been compromised in MDR E. faecalis. In restriction-modification (R-M) defense, the bacterial genome is methylated at cytosine (C) or adenine (A) residues by a methyltransferase (MTase), such that nonself DNA can be distinguished from self DNA. A cognate restriction endonuclease digests improperly modified nonself DNA. Little is known about R-M in E. faecalis. Here, we use genome resequencing to identify DNA modifications occurring in the oral isolate OG1RF. OG1RF has one of the smallest E. faecalis genomes sequenced to date and possesses few MGEs. Single-molecule real-time (SMRT) and bisulfite sequencing revealed that OG1RF has global 5-methylcytosine (m5C) methylation at 5=-GC-WGC-3= motifs. A type II R-M system confers the m5C modification, and disruption of this system impacts OG1RF electrotransformability and conjugative transfer of an antibiotic resistance plasmid. A second DNA MTase was poorly expressed under laboratory conditions but conferred global N 4 -methylcytosine (m4C) methylation at 5=-CCGG-3= motifs when expressed in Escherichia coli. Based on our results, we conclude that R-M can act as a barrier to MGE acquisition and likely influences antibiotic resistance gene dissemination in the E. faecalis species. IMPORTANCEThe horizontal transfer of antibiotic resistance genes among bacteria is a critical public health concern. Enterococcus faecalis is an opportunistic pathogen that causes life-threatening infections in humans. Multidrug resistance acquired by horizontal gene transfer limits treatment options for these infections. In this study, we used innovative DNA sequencing methodologies to investigate how a model strain of E. faecalis discriminates its own DNA from foreign DNA, i.e., self versus nonself discrimination. We also assess the role of an E. faecalis genome modification system in modulating conjugative transfer of an antibiotic resistance plasmid. These results are significant because they demonstrate that differential genome modification impacts horizontal gene transfer frequencies in E. faecalis. Enterococcus faecalis is a Gram-positive bacterium that natively colonizes the gastrointestinal tracts of humans and other animals (1). It is an opportunistic pathogen that causes life-threatening infections, such as bacteremia and endocarditis in compromised individuals (2). E. faecalis is among the leading causes of hospital-acquired infections in the United States, making it a major public health concern (3). Rising antibiotic resistance...

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Section: Resultsmentioning

confidence: 99%

Genome Modification in Enterococcus faecalis OG1RF Assessed by Bisulfite Sequencing and Single-Molecule Real-Time Sequencing

et al. 2015

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“…Inclusion of 10% CL in model membranes causes more avid membrane binding of daptomycin, as evidenced by both ITC and monolayer compression experiments. 66 However, in the liposome model at least, CL inhibits membrane permeabilization, 66 which suggests that it might mediate resistance to daptomycin by competing for it with PG. Such an effect has indeed been claimed to occur in vivo, 67 but the CL-enriched membrane domains purported in that study were identified by staining with nonyl-acridine orange, which has since been shown to bind to phosphatidylglycerol also.…”

Section: Lipid Specificitymentioning

confidence: 99%

“…Nevertheless, some useful insights have been obtained concerning the distribution of daptomycin Top and side view of the octamer that spans both leaflets and forms a functional pore. C: A tetrameric form that is confined to the outer leaflet has been observed on membranes containing cardiolipin 66 and may also occur at sub-saturating calcium concentrations. 84 across the two membrane leaflets, the positions of individual residues relative to the bilayer, and the mutual proximity of residues within the oligomer.…”

Section: Topology Of Membrane-associated Daptomycinmentioning

confidence: 99%

“…From these findings, the hypothetical octameric structure shown in Figure 6 was proposed. 66 Various fluorescence studies have identified membrane-inserted residues. The intrinsic fluorescence of Kyn (13) undergoes a strong blue-shift and increase in quantum yield.…”

Section: Topology Of Membrane-associated Daptomycinmentioning

confidence: 99%

“…Increased lipid flip-flop was demonstrated with a lipid analogue that incorporates the somewhat polar NBD label into a shorted acyl side chain, 42 but not with a head group-labelled lipid probe. 66 Both lipid analogues are synthetic, and thus the relevance of both of these observations to natural bacterial membranes remains uncertain.…”

Section: Membrane Fusion Flip-flop and Lipid Extractionmentioning

confidence: 99%

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The action mechanism of daptomycin

Taylor

Palmer

2016

Bioorganic & Medicinal Chemistry

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223

172

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Daptomycin is a lipopeptide antibiotic produced by the soil bacterium Streptomyces roseosporus that is clinically used to treat severe infections with Grampositive bacteria. In this review, we discuss the mode of action of this important antibiotic. Although daptomycin is structurally related to amphomycin and similar lipopeptides that inhibit peptidoglycan biosynthesis, experimental studies have not produced clear evidence that daptomycin shares their action mechanism. Instead, the best characterized effect of daptomycin is the permeabilization and depolarization of the bacterial cell membrane. This activity, which can account for daptomycin's bactericidal effect, correlates with the level of phosphatidylglycerol (PG) in the membrane. Accordingly, reduced synthesis of PG or its increased conversion to lysyl-PG promotes bacterial resistance to daptomycin. While other resistance mechanisms suggest that daptomycin may indeed directly interfere with cell wall synthesis or cell division, such effects still await direct experimental confirmation. Daptomycin's complex structure and biosynthesis have hampered the analysis of its structure activity relationships. Novel methods of total synthesis, including a recent one that is carried out entirely on a solid phase, will enable a more thorough and systematic exploration of the sequence space.

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Cardiolipin prevents pore formation in phosphatidylglycerol bacterial membrane models

et al. 2021

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Several antimicrobial peptides, including magainin and the human cathelicidin LL‐37, act by forming pores in bacterial membranes. Bacteria such as Staphylococcus aureus modify their membrane's cardiolipin composition to resist such types of perturbations that compromise their membrane stability. Here, we used molecular dynamic simulations to quantify the role of cardiolipin on the formation of pores in simple bacterial‐like membrane models composed of phosphatidylglycerol and cardiolipin mixtures. Cardiolipin modified the structure and ordering of the lipid bilayer, making it less susceptible to mechanical changes. Accordingly, the free‐energy barrier for the formation of a transmembrane pore and its kinetic instability augmented by increasing the cardiolipin concentration. This is attributed to the unfavorable positioning of cardiolipin near the formed pore, due to its small polar head and bulky hydrophobic body. Overall, our study demonstrates how cardiolipin prevents membrane‐pore formation and this constitutes a plausible mechanism used by bacteria to act against stress perturbations and, thereby, gain resistance to antimicrobial agents.

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Cardiolipin Prevents Membrane Translocation and Permeabilization by Daptomycin

Cited by 141 publications

References 27 publications

Genome Modification in Enterococcus faecalis OG1RF Assessed by Bisulfite Sequencing and Single-Molecule Real-Time Sequencing

Genome Modification in Enterococcus faecalis OG1RF Assessed by Bisulfite Sequencing and Single-Molecule Real-Time Sequencing

The action mechanism of daptomycin

Cardiolipin prevents pore formation in phosphatidylglycerol bacterial membrane models

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