Host-selected mutations converging on a global regulator drive an adaptive leap by bacteria to symbiosis

Pankey, M. Sabrina; Foxall, Randi L; Ster, Ian M; Perry, Lauren A; Schuster, Brian G.; Donner, Rachel A; Coyle, Matthew; Cooper, Vaughn S.; Whistler, Cheryl A.

doi:10.1101/067025

Cited by 3 publications

(4 citation statements)

References 132 publications

(174 reference statements)

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“…6). The effect of binK disruption on syp and bcs transcription is consistent with recent reports (33,34). These data suggest that (i) Calcium-dependent cell clumping depends on sypF and sypG.…”

Section: Nt Of Kv7949 With Ckv8297supporting

confidence: 91%

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Discovery of Calcium as a Biofilm-Promoting Signal for Vibrio fischeri Reveals New Phenotypes and Underlying Regulatory Complexity

et al. 2018

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uses biofilm formation to promote symbiotic colonization of its squid host, Control over biofilm formation is exerted at the level of transcription of the symbiosis polysaccharide () locus by a complex set of two-component regulators. Biofilm formation can be induced by overproduction of the sensor kinase RscS, which requires the activities of the hybrid sensor kinase SypF and the response regulator SypG and is negatively regulated by the sensor kinase BinK. Here, we identify calcium as a signal that promotes biofilm formation by biofilm-competent strains under conditions in which biofilms are not typically observed (growth with shaking). This was true for RscS-overproducing cells as well as for strains in which only the negative regulator was deleted. The latter results provided, for the first time, an opportunity to induce and evaluate biofilm formation without regulator overexpression. Using these conditions, we determined that calcium induces both-dependent and bacterial cellulose synthesis ()-dependent biofilms at the level of transcription of these loci. The calcium-induced biofilms were dependent on SypF, but SypF's Hpt domain was sufficient for biofilm formation. These data suggested the involvement of another sensor kinase(s) and led to the discovery that both RscS and a previously uncharacterized sensor kinase, HahK, functioned in this pathway. Together, the data presented here reveal both a new signal and biofilm phenotype produced by cells, the coordinate production of two polysaccharides involved in distinct biofilm behaviors, and a new regulator that contributes to control over these processes. Biofilms, or communities of surface-attached microorganisms adherent via a matrix that typically includes polysaccharides, are highly resistant to environmental stresses and are thus problematic in the clinic and important to study. forms biofilms to colonize its symbiotic host, making this organism useful for studying biofilms. Biofilm formation depends on the polysaccharide locus and its regulators. Here, we identify a signal, calcium, that induces both SYP-PS and cellulose-dependent biofilms. We also identify a new regulator, the sensor kinase HahK, and discover a mutant phenotype for the sensor kinase RscS. This work thus reveals a specific biofilm-inducing signal that coordinately controls two polysaccharides, identifies a new regulator, and clarifies the regulatory control over biofilm formation by.

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Section: Nt Of Kv7949 With Ckv8297supporting

confidence: 91%

“…Recently, the involvement of a third sensor kinase, BinK, was reported (33,34). BinK inhibits the production of syp-dependent biofilms induced by RscS overexpression, and loss of BinK enhances symbiotic biofilm formation and colonization ( Fig.…”

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confidence: 99%

Discovery of Calcium as a Biofilm-Promoting Signal for Vibrio fischeri Reveals New Phenotypes and Underlying Regulatory Complexity

et al. 2018

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“…5, Supplementary Table 7, Supplementary Table 8). In detail, biofilm formation can enable persistent contact with the host and increase stress-tolerance 13,14 , exemplified by many pathogens 15 , thereby improving survival in the nematode's digestive tract. As a consequence of increased biofilm formation, aggregated cells may be expelled more easily 16 , thereby explaining the observed increase in release.…”

Section: Evolution Of Host Specialist Bacteriamentioning

confidence: 99%

Bacterial c-di-GMP plays a key role in the evolution of host-association

Obeng

Czerwinski

Schuetz

et al. 2023

Preprint

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Most microbes evolve faster than their hosts and should therefore drive evolution of host-microbe interactions. However, relatively little is known about the characteristics that define the adaptive path of microbes to host-association. In this study we have identified microbial traits that mediate adaptation to hosts by experimentally evolving the bacterium Pseudomonas lurida with the nematode Caenorhabditis elegans. We repeatedly observed the evolution of beneficial host-specialist bacteria with improved persistence in the nematode, achieved by mutations that uniformly upregulate the universal second messenger c-di-GMP. We subsequently upregulated c-di-GMP in different Pseudomonas species, consistently causing increased host-association. Comparison of Pseudomonad genomes from various environments revealed that c-di-GMP underlies adaptation to a variety of hosts, from plants to humans, suggesting that it is fundamental for establishing host-association.

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“…approaches (7,29), thus increasing the need for tools and approaches to facilitate manipulations of V. fischeri.…”

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confidence: 99%

Tools for Rapid Genetic Engineering of Vibrio fischeri

Visick

Hodge‐Hanson

Tischler

et al. 2018

Appl Environ Microbiol

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is used as a model for a number of processes, including symbiosis, quorum sensing, bioluminescence, and biofilm formation. Many of these studies depend on generating deletion mutants and complementing them. Engineering such strains, however, is a time-consuming, multistep process that relies on cloning and subcloning. Here, we describe a set of tools that can be used to rapidly engineer deletions and insertions in the chromosome without cloning. We developed a uniform approach for generating deletions using PCR splicing by overlap extension (SOEing) with antibiotic cassettes flanked by standardized linker sequences. PCR SOEing of the cassettes to sequences up- and downstream of the target gene generates a DNA product that can be directly introduced by natural transformation. Selection for the introduced antibiotic resistance marker yields the deletion of interest in a single step. Because these cassettes also contain FRT (FLP recognition target) sequences flanking the resistance marker, Flp recombinase can be used to generate an unmarked, in-frame deletion. We developed a similar methodology and tools for the rapid insertion of specific genes at a benign site in the chromosome for purposes such as complementation. Finally, we generated derivatives of these tools to facilitate different applications, such as inducible gene expression and assessing protein production. We demonstrated the utility of these tools by deleting and inserting genes known or predicted to be involved in motility. While developed for strain ES114, we anticipate that these tools can be adapted for use in other strains and, potentially, other microbes. is a model organism for studying a variety of important processes, including symbiosis, biofilm formation, and quorum sensing. To facilitate investigation of these biological mechanisms, we developed approaches for rapidly generating deletions and insertions and demonstrated their utility using two genes of interest. The ease, consistency, and speed of the engineering is facilitated by a set of antibiotic resistance cassettes with common linker sequences that can be amplified by PCR with universal primers and fused to adjacent sequences using splicing by overlap extension and then introduced directly into , eliminating the need for cloning and plasmid conjugation. The antibiotic cassettes are flanked by FRT sequences, permitting their removal using Flp recombinase. We augmented these basic tools with a family of constructs for different applications. We anticipate that these tools will greatly accelerate mechanistic studies of biological processes in and potentially other species.

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Host-selected mutations converging on a global regulator drive an adaptive leap by bacteria to symbiosis

Cited by 3 publications

References 132 publications

Discovery of Calcium as a Biofilm-Promoting Signal for Vibrio fischeri Reveals New Phenotypes and Underlying Regulatory Complexity

Discovery of Calcium as a Biofilm-Promoting Signal for Vibrio fischeri Reveals New Phenotypes and Underlying Regulatory Complexity

Bacterial c-di-GMP plays a key role in the evolution of host-association

Tools for Rapid Genetic Engineering of Vibrio fischeri

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