In Situ Structure of an Intact Lipopolysaccharide-Bound Bacterial Surface Layer

Kügelgen, Andriko von; Tang, Haiping; Hardy, Gail G.; Kureisaite-Ciziene, Danguole; Brun, Yves V.; Stansfeld, Phillip J.; Robinson, Carol V.; Bharat, Tanmay A. M.

doi:10.1016/j.cell.2019.12.006

Cited by 98 publications

(114 citation statements)

References 70 publications

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“…5E and Movie S5) or lined up in close proximity to the outer membrane, separated only by lipopolysaccharide and other cell surface molecules, that are known to be present in the outer membrane of Gram-negative bacteria including P. aeruginosa (SI Appendix, Fig. S8I) at high copy numbers (24,25). These cryo-ET images provide snapshots of the association of Pf4 liquid crystalline droplets and the P. aeruginosa cell surface, indicating that Pf4 filaments can deform and wrap around rod-shaped bacterial cells to begin encapsulation.…”

Section: P Aeruginosa Antibiotic Protection Mediated By Pf4 Liquid Cmentioning

confidence: 99%

Phage liquid crystalline droplets form occlusive sheaths that encapsulate and protect infectious rod-shaped bacteria

Tarafder

Kügelgen

Mellul

et al. 2020

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

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137

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The opportunistic pathogenPseudomonas aeruginosais a major cause of antibiotic-tolerant infections in humans.P. aeruginosaevades antibiotics in bacterial biofilms by up-regulating expression of a symbiotic filamentous inoviral prophage, Pf4. We investigated the mechanism of phage-mediated antibiotic tolerance using biochemical reconstitution combined with structural biology and high-resolution cellular imaging. We resolved electron cryomicroscopy atomic structures of Pf4 with and without its linear single-stranded DNA genome, and studied Pf4 assembly into liquid crystalline droplets using optical microscopy and electron cryotomography. By biochemically replicating conditions necessary for antibiotic protection, we found that phage liquid crystalline droplets form phase-separated occlusive compartments around rod-shaped bacteria leading to increased bacterial survival. Encapsulation by these compartments was observed even when inanimate colloidal rods were used to mimic rod-shaped bacteria, suggesting that shape and size complementarity profoundly influences the process. Filamentous inoviruses are pervasive across prokaryotes, and in particular, several Gram-negative bacterial pathogens includingNeisseria meningitidis,Vibrio cholerae,andSalmonella entericaharbor these prophages. We propose that biophysical occlusion mediated by secreted filamentous molecules such as Pf4 may be a general strategy of bacterial survival in harsh environments.

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Section: P Aeruginosa Antibiotic Protection Mediated By Pf4 Liquid Cmentioning

confidence: 99%

Phage liquid crystalline droplets form occlusive sheaths that encapsulate and protect infectious rod-shaped bacteria

Tarafder

Kügelgen

Mellul

et al. 2020

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

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137

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“…Cryo-electron tomography (cryo-ET) combined with image processing approaches, such as subtomogram averaging (STA) can provide structural insights into native cellular environments 17 . Medium to high-resolution structures of proteins within their native environment using these methods have so far been only described for a few specimens, often large or highly symmetrical assemblies [18][19][20][21][22] . Among the limitations that restrict the resolution that can be obtained in cells are the thickness of the specimen, reduction of which often requires additional specimen preparation steps, and the number of protein complexes available for generating a higher-resolution structure 17 .…”

mentioning

confidence: 99%

Cryo-electron tomography structure of Arp2/3 complex in cells reveals new insights into the branch junction

et al. 2020

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The actin-related protein (Arp)2/3 complex nucleates branched actin filament networks pivotal for cell migration, endocytosis and pathogen infection. Its activation is tightly regulated and involves complex structural rearrangements and actin filament binding, which are yet to be understood. Here, we report a 9.0 Å resolution structure of the actin filament Arp2/3 complex branch junction in cells using cryo-electron tomography and subtomogram averaging. This allows us to generate an accurate model of the active Arp2/3 complex in the branch junction and its interaction with actin filaments. Notably, our model reveals a previously undescribed set of interactions of the Arp2/3 complex with the mother filament, significantly different to the previous branch junction model. Our structure also indicates a central role for the ArpC3 subunit in stabilizing the active conformation.

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“…A), rotatable images (e.g. C), and realistic graphics coupled with high‐resolution imaging to demonstrate relative position and very fine detail from nanometre to micrometre scales (brilliantly exemplified by using cryo‐electron microscopy to illustrate cell surface structures in Caulobacter crescentus by von Kügelgen et al (2019)), all add an extra dimension that can help to attract children to the wonder of microbes, their structures, activities and interactions. Goodsell et al (2020) review the plethora of techniques that are allowing cellular machines, compartments and macromolecules to be visualized, and they consider the important issue of the use of artistic licence, particularly to stimulate interest and to educate the non‐technical audience.…”

Section: Discussionmentioning

confidence: 99%

Visualizing the invisible: class excursions to ignite children’s enthusiasm for microbes

McGenity

Gessesse

Hallsworth

et al. 2020

Microbial Biotechnology

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Summary We have recently argued that, because microbes have pervasive – often vital – influences on our lives, and that therefore their roles must be taken into account in many of the decisions we face, society must become microbiology‐literate, through the introduction of relevant microbiology topics in school curricula (Timmis et al. 2019. Environ Microbiol 21: 1513‐1528). The current coronavirus pandemic is a stark example of why microbiology literacy is such a crucial enabler of informed policy decisions, particularly those involving preparedness of public‐health systems for disease outbreaks and pandemics. However, a significant barrier to attaining widespread appreciation of microbial contributions to our well‐being and that of the planet is the fact that microbes are seldom visible: most people are only peripherally aware of them, except when they fall ill with an infection. And it is disease, rather than all of the positive activities mediated by microbes, that colours public perception of ‘germs’ and endows them with their poor image. It is imperative to render microbes visible, to give them life and form for children (and adults), and to counter prevalent misconceptions, through exposure to imagination‐capturing images of microbes and examples of their beneficial outputs, accompanied by a balanced narrative. This will engender automatic mental associations between everyday information inputs, as well as visual, olfactory and tactile experiences, on the one hand, and the responsible microbes/microbial communities, on the other hand. Such associations, in turn, will promote awareness of microbes and of the many positive and vital consequences of their actions, and facilitate and encourage incorporation of such consequences into relevant decision‐making processes. While teaching microbiology topics in primary and secondary school is key to this objective, a strategic programme to expose children directly and personally to natural and managed microbial processes, and the results of their actions, through carefully planned class excursions to local venues, can be instrumental in bringing microbes to life for children and, collaterally, their families. In order to encourage the embedding of microbiology‐centric class excursions in current curricula, we suggest and illustrate here some possibilities relating to the topics of food (a favourite pre‐occupation of most children), agriculture (together with horticulture and aquaculture), health and medicine, the environment and biotechnology. And, although not all of the microbially relevant infrastructure will be within reach of schools, there is usually access to a market, local food store, wastewater treatment plant, farm, surface water body, etc., all of which can provide opportunities to explore microbiology in action. If children sometimes consider the present to be mundane, even boring, they are usually excited with both the past and the future so, where possible, visits to local museums (the past) and research institutions advancing knowledge frontiers (the...

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In Situ Structure of an Intact Lipopolysaccharide-Bound Bacterial Surface Layer

Cited by 98 publications

References 70 publications

Phage liquid crystalline droplets form occlusive sheaths that encapsulate and protect infectious rod-shaped bacteria

Phage liquid crystalline droplets form occlusive sheaths that encapsulate and protect infectious rod-shaped bacteria

Cryo-electron tomography structure of Arp2/3 complex in cells reveals new insights into the branch junction

Visualizing the invisible: class excursions to ignite children’s enthusiasm for microbes

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