Dataset Information

Persister Cells - a Plausible Outcome of Neutral Coevolutionary Drift.

ABSTRACT: The phenomenon of bacterial persistence - a non-inherited antibiotic tolerance in a minute fraction of the bacterial population, was observed more than 70 years ago. Nowadays, it is suggested that "persister cells" undergo an alternative scenario of the cell cycle; however, pathways involved in its emergence are still not identified. We present a mathematically grounded scenario of such possibility. We have determined that population drift in the space of multiple neutrally coupled mutations, which we called "neutrally coupled co-evolution" (NCCE), leads to increased dynamic complexity of bacterial populations via appearance of cells capable of carrying out a single cell cycle in two or more alternative ways and that universal properties of the coupled transcription-translation system underlie this phenotypic multiplicity. According to our hypothesis, modern persister cells have derived from such cells and regulatory mechanisms that govern the consolidation of this phenomenon represented the trigger. We assume that the described type of neutrally coupled co-evolution could play an important role in the origin of extremophiles, both in bacteria and archaea.

SUBMITTER: Khlebodarova TM

PROVIDER: S-EPMC6156226 | biostudies-literature | 2018 Sep

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Persister Cells - a Plausible Outcome of Neutral Coevolutionary Drift.

Khlebodarova T M TM Likhoshvai V A VA

Scientific reports 20180925 1

The phenomenon of bacterial persistence - a non-inherited antibiotic tolerance in a minute fraction of the bacterial population, was observed more than 70 years ago. Nowadays, it is suggested that "persister cells" undergo an alternative scenario of the cell cycle; however, pathways involved in its emergence are still not identified. We present a mathematically grounded scenario of such possibility. We have determined that population drift in the space of multiple neutrally coupled mutations, wh ...[more]

PMID: 30254316

Similar Datasets

Project description:Unspecific peroxygenase (UPO) is a highly promiscuous biocatalyst, and its selective mono(per)oxygenase activity makes it useful for many synthetic chemistry applications. Among the broad repertory of library creation methods for directed enzyme evolution, genetic drift allows neutral mutations to be accumulated gradually within a polymorphic network of variants. In this study, we conducted a campaign of genetic drift with UPO in Saccharomyces cerevisiae, so that neutral mutations were simply added and recombined in vivo With low mutational loading and an activity threshold of 45% of the parent's native function, mutant libraries enriched in folded active UPO variants were generated. After only eight rounds of genetic drift and DNA shuffling, we identified an ensemble of 25 neutrally evolved variants with changes in peroxidative and peroxygenative activities, kinetic thermostability, and enhanced tolerance to organic solvents. With an average of 4.6 substitutions introduced per clone, neutral mutations covered approximately 10% of the protein sequence. Accordingly, this study opens new avenues for UPO design by bringing together neutral genetic drift and DNA recombination in vivoIMPORTANCE Fungal peroxygenases resemble the peroxide shunt pathway of cytochrome P450 monoxygenases, performing selective oxyfunctionalizations of unactivated C-H bonds in a broad range of organic compounds. In this study, we combined neutral genetic drift and in vivo DNA shuffling to generate highly functional peroxygenase mutant libraries. The panel of neutrally evolved peroxygenases showed different activity profiles for peroxygenative substrates and improved stability with respect to temperature and the presence of organic cosolvents, making the enzymes valuable blueprints for emerging evolution campaigns. This association of DNA recombination and neutral drift is paving the way for future work in peroxygenase engineering and, from a more general perspective, to any other enzyme system heterologously expressed in S. cerevisiae.

Project description:Bacterial persistence is a feature that allows susceptible bacteria to survive extreme concentrations of antibiotics and it has been verified in a number of species, such as Escherichia coli, Pseudomonas aeruginosa, Staphylococcus spp., Mycobacterium spp. However, even though Acinetobacter baumannii is an important nosocomial pathogen, data regarding its persistence phenotype are still lacking. Therefore, the aim of this study was to evaluate the persistence phenotype in A. baumannii strains, as well as its variation among strains after treatment with polymyxin B and tobramycin. Stationary cultures of 37 polymyxin B-susceptible clinical strains of A. baumannii were analyzed for surviving cells after exposure to 15 µg/mL of polymyxin B for 6 h, by serial dilutions and colony counting. Among these, the 30 tobramycin-susceptible isolates also underwent tobramycin treatment at a concentration of 160 µg/mL and persister cells occurrence was evaluated equally. A high heterogeneity of persister cells formation patterns among isolates was observed. Polymyxin B-treated cultures presented persister cells corresponding from 0.0007% to 10.1% of the initial population and two isolates failed to produce detectable persister cells under this condition. A high variability could also be observed when cells were treated with tobramycin: the persister fraction corresponded to 0.0003%-11.84% of the pre-treatment population. Moreover, no correlation was found between persister subpopulations comparing both antibiotics among isolates, indicating that different mechanisms underlie the internal control of this phenotype. This is the first report of persister cells occurrence in A. baumannii. Our data suggest that distinct factors regulate the tolerance for unrelated antibiotics in this species, contrasting the multi-drug tolerance observed in other species (eg. dormancy-mediated tolerance). Supporting this observation, polymyxin B--an antibiotic that is believed to act on non-dividing cells as well--failed to eradicate persister cells in the majority of the isolates, possibly reflecting a disconnection between persistence and dormancy.

Dataset Information

Persister Cells - a Plausible Outcome of Neutral Coevolutionary Drift.

Publications

Persister Cells - a Plausible Outcome of Neutral Coevolutionary Drift.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets