Two enhancer binding proteins activate σ54-dependent transcription of a quorum regulatory RNA in a bacterial symbiont

Surrett, Ericka D; Guckes, Kirsten R.; Cousins, Shyan; Ruskoski, Terry B; Cecere, Andrew G.; Ludvik, Denise A; Okafor, C. Denise; Mandel, Mark J.; Miyashiro, Tim

doi:10.7554/elife.78544

Cited by 4 publications

(1 citation statement)

References 59 publications

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“…For strains that have not been previously evaluated by this method, we suggest using 10-fold dilutions that range from 10 1 −10 4 CFU/mL to obtain an estimate for the SD 50 value. We also note that the inoculum duration used here (3.5 h) is commonly used by us and others to study ES114 because it is sufficiently long for cells to form the aggregates that promote host colonization [ 24 , 25 ]. The duration does represent a variable that can be changed, but the resulting SD 50 values would likely be affected.…”

Section: Expected Resultsmentioning

confidence: 99%

Quantification of the capacity of vibrio fischeri to establish symbiosis with Euprymna scolopes

Donnelly,

Giacobe,

Cook

et al. 2023

PLoS ONE

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Most animals establish long-term symbiotic associations with bacteria that are critical for normal host physiology. The symbiosis that forms between the Hawaiian squid Euprymna scolopes and the bioluminescent bacterium Vibrio fischeri serves as an important model system for investigating the molecular mechanisms that promote animal-bacterial symbioses. E. scolopes hatch from their eggs uncolonized, which has led to the development of squid-colonization assays that are based on introducing culture-grown V. fischeri cells to freshly hatched juvenile squid. Recent studies have revealed that strains often exhibit large differences in how they establish symbiosis. Therefore, we sought to develop a simplified and reproducible protocol that permits researchers to determine appropriate inoculum levels and provides a platform to standardize the assay across different laboratories. In our protocol, we adapt a method commonly used for evaluating the infectivity of pathogens to quantify the symbiotic capacity of V. fischeri strains. The resulting metric, the symbiotic dose-50 (SD50), estimates the inoculum level that is necessary for a specific V. fischeri strain to establish a light-emitting symbiosis. Relative to other protocols, our method requires 2–5-fold fewer animals. Furthermore, the power analysis presented here suggests that the protocol can detect up to a 3-fold change in the SD50 between different strains.

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Section: Expected Resultsmentioning

confidence: 99%

Quantification of the capacity of vibrio fischeri to establish symbiosis with Euprymna scolopes

Donnelly,

Giacobe,

Cook

et al. 2023

PLoS ONE

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Quorum sensing inhibits interference competition among bacterial symbionts within a host

Guckes,

Yount,

Steingard

et al. 2023

Current Biology

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Vibrio fischeri : a model for host-associated biofilm formation

Fung,

Esin,

Visick

2024

J Bacteriol

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Multicellular communities of adherent bacteria known as biofilms are often detrimental in the context of a human host, making it important to study their formation and dispersal, especially in animal models. One such model is the symbiosis between the squid Euprymna scolopes and the bacterium Vibrio fischeri . Juvenile squid hatch aposymbiotically and selectively acquire their symbiont from natural seawater containing diverse environmental microbes. Successful pairing is facilitated by ciliary movements that direct bacteria to quiet zones on the surface of the squid’s symbiotic light organ where V. fischeri forms a small aggregate or biofilm. Subsequently, the bacteria disperse from that aggregate to enter the organ, ultimately reaching and colonizing deep crypt spaces. Although transient, aggregate formation is critical for optimal colonization and is tightly controlled. In vitro studies have identified a variety of polysaccharides and proteins that comprise the extracellular matrix. Some of the most well-characterized matrix factors include the symbiosis polysaccharide (SYP), cellulose polysaccharide, and LapV adhesin. In this review, we discuss these components, their regulation, and other less understood V. fischeri biofilm contributors. We also highlight what is currently known about dispersal from these aggregates and host cues that may promote it. Finally, we briefly describe discoveries gleaned from the study of other V. fischeri isolates. By unraveling the complexities involved in V. fischeri ’s control over matrix components, we may begin to understand how the host environment triggers transient biofilm formation and dispersal to promote this unique symbiotic relationship.

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Two enhancer binding proteins activate σ54-dependent transcription of a quorum regulatory RNA in a bacterial symbiont

Cited by 4 publications

References 59 publications

Quantification of the capacity of vibrio fischeri to establish symbiosis with Euprymna scolopes

Quantification of the capacity of vibrio fischeri to establish symbiosis with Euprymna scolopes

Quorum sensing inhibits interference competition among bacterial symbionts within a host

Vibrio fischeri : a model for host-associated biofilm formation

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