Minicells as a Damage Disposal Mechanism in Escherichia coli

Rang, Camilla U.; Proenca, Audrey Menegaz; Buetz, Christen; Shi, Chao; Chao, Lin

doi:10.1128/msphere.00428-18

Cited by 36 publications

(31 citation statements)

References 78 publications

(90 reference statements)

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“…To visualize the biasing of damage loads toward old daughters in our microfluidic device, we cultured E. coli expressing the small chaperone inclusion body protein A (IbpA) fusioned to yellow fluorescent protein (YFP). IbpA-YFP was shown to colocalize with protein aggregates in bacterial cells [11], thus serving as a marker for the presence and position of nongenetic damage [24]. By culturing this strain in our microfluidic device, we observed the progressive accumulation of damage in the old poles of lineages in a state of equilibrium ( Fig 1F-1M).…”

Section: Large Protein Aggregates Become Anchored At Old Cell Polesmentioning

confidence: 85%

“…Experiments were performed with K-12 E. coli wild-type strain MG1655 for the determination of damage accumulation in immortally proliferating bacteria and for experiments on the disruption of growth equilibrium. The visualization of protein aggregates was performed with MG1655 E. coli expressing YFP bound to the small heat-shock protein IbpA, constructed according to Rang and colleagues [24] from the construct IbpA-yfp-Cm r kindly provided by Ariel B. Lindner (INSERM, France) [11]. Repair mutants were screened for asymmetry and equilibrium using E. coli BW25113 ΔclpB (CGSC #11763) and ΔdnaK (CGSC #8342) from the Keio knockout collection [35].…”

Section: Bacterial Strains and Growth Conditionsmentioning

confidence: 99%

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Cell aging preserves cellular immortality in the presence of lethal levels of damage

et al. 2019

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Cellular aging, a progressive functional decline driven by damage accumulation, often culminates in the mortality of a cell lineage. Certain lineages, however, are able to sustain longlasting immortality, as prominently exemplified by stem cells. Here, we show that Escherichia coli cell lineages exhibit comparable patterns of mortality and immortality. Through single-cell microscopy and microfluidic techniques, we find that these patterns are explained by the dynamics of damage accumulation and asymmetric partitioning between daughter cells. At low damage accumulation rates, both aging and rejuvenating lineages retain immortality by reaching their respective states of physiological equilibrium. We show that both asymmetry and equilibrium are present in repair mutants lacking certain repair chaperones, suggesting that intact repair capacity is not essential for immortal proliferation. We show that this growth equilibrium, however, is displaced by extrinsic damage in a dosagedependent response. Moreover, we demonstrate that aging lineages become mortal when damage accumulation rates surpass a threshold, whereas rejuvenating lineages within the same population remain immortal. Thus, the processes of damage accumulation and partitioning through asymmetric cell division are essential in the determination of proliferative mortality and immortality in bacterial populations. This study provides further evidence for the characterization of cellular aging as a general process, affecting prokaryotes and eukaryotes alike and according to similar evolutionary constraints.

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Section: Large Protein Aggregates Become Anchored At Old Cell Polesmentioning

confidence: 85%

Section: Bacterial Strains and Growth Conditionsmentioning

confidence: 99%

Cell aging preserves cellular immortality in the presence of lethal levels of damage

et al. 2019

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“…Another example is the cell division protein MinD, a thermo-labile protein that is markedly destabilized by elevated KJE levels, even at NC. The inhibition of MinD function causes minicell formation, which was proposed to facilitate the disposal of damaged proteins under stress conditions (de Boer et al, 1989;Rang et al, 2018), suggesting that destabilization by KJE may be physiologically relevant. As MinD consists of a single c.37 domain, these findings show that the chaperone machinery can have opposing effects on protein stability, even for proteins with similar c.37 domain topologies.…”

Section: Destabilizing Effects Of Kjementioning

confidence: 99%

The Hsp70 Chaperone System Stabilizes a Thermo-sensitive Subproteome in E. coli

et al. 2019

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Highlights d Acute heat stress causes unfolding of a thermo-sensitive subproteome in E. coli d DnaK (Hsp70) markedly stabilizes most thermo-sensitive proteins d Numerous essential ribosomal proteins are conformationally stabilized by DnaK d Other DnaK-stabilized proteins tend to be large, multidomain, and hetero-oligomeric SUMMARYStress-inducible molecular chaperones have essential roles in maintaining protein homeostasis, but the extent to which they affect overall proteome stability remains unclear. Here, we analyze the effects of the DnaK (Hsp70) system on protein stability in Escherichia coli using pulse proteolysis combined with quantitative proteomics. We quantify $1,500 soluble proteins and find $500 of these to be protease sensitive under normal growth conditions, indicating a high prevalence of conformationally dynamic proteins, forming a metastable subproteome. Acute heat stress results in the unfolding of an additional $200 proteins, reflected in the exposure of otherwise buried hydrophobic regions. Overexpression of the DnaK chaperone system markedly stabilizes numerous thermo-sensitive proteins, including multiple ribosomal proteins and large, hetero-oligomeric proteins containing the evolutionarily ancient c.37 fold (P loop nucleoside triphosphate hydrolases). Thus, the Hsp70 system, in addition to its known chaperone functions, has a remarkable capacity to stabilize proteins in their folded states under denaturing stress conditions.

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“…Both Gram-positive and Gram-negative bacteria have the ability to form minicells but little is known about why this happens. A recent study [24] suggests that minicells could act as a "damage disposal" mechanism for proteins damaged by antibiotics.…”

Section: Angewandte Chemiementioning

confidence: 99%

DNA Nanostructures for Targeted Antimicrobial Delivery

et al. 2020

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We report the use of DNA origami nanostructures, functionalized with aptamers, as a vehicle for delivering the antibacterial enzyme lysozyme in a specific and efficient manner. We test the system against Gram‐positive (Bacillus subtilis) and Gram‐negative (Escherichia coli) targets. We use direct stochastic optical reconstruction microscopy (dSTORM) and atomic force microscopy (AFM) to characterize the DNA origami nanostructures and structured illumination microscopy (SIM) to assess the binding of the origami to the bacteria. We show that treatment with lysozyme‐functionalized origami slows bacterial growth more effectively than treatment with free lysozyme. Our study introduces DNA origami as a tool in the fight against antibiotic resistance, and our results demonstrate the specificity and efficiency of the nanostructure as a drug delivery vehicle.

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Minicells as a Damage Disposal Mechanism in Escherichia coli

Cited by 36 publications

References 78 publications

Cell aging preserves cellular immortality in the presence of lethal levels of damage

Cell aging preserves cellular immortality in the presence of lethal levels of damage

The Hsp70 Chaperone System Stabilizes a Thermo-sensitive Subproteome in E. coli

DNA Nanostructures for Targeted Antimicrobial Delivery

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