Meddling with Metal Sensors: Fur-Family Proteins as Signaling Hubs.

The ferric uptake regulator (Fur) protein is the founding member of the FUR superfamily of metalloregulatory proteins that control metal homeostasis in bacteria. FUR proteins regulate metal homeostasis in response to the binding of iron (Fur), zinc (Zur), manganese (Mur), or nickel (Nur). FUR family proteins are generally dimers in solution, but the DNA-bound complex can involve a single dimer, a dimer-of-dimers, or an extended array of bound protein. Elevated FUR levels due to changes in cell physiology increase DNA occupancy and may also kinetically facilitate protein dissociation. Interactions between FUR proteins and other regulators are commonplace, often including cooperative and competitive DNA-binding interactions within the regulatory region. Further, there are many emerging examples of allosteric regulators that interact directly with FUR family proteins. Here, we focus on newly uncovered examples of allosteric regulation by diverse Fur antagonists (Escherichia coli YdiV/SlyD, Salmonella enterica EIIANtr, Vibrio parahaemolyticus FcrX, Acinetobacter baumannii BlsA, Bacillus subtilis YlaN, and Pseudomonas aeruginosa PacT) as well as one Zur antagonist (Mycobacterium bovis CmtR). Small molecules and metal complexes may also serve as regulatory ligands, with examples including heme binding to Bradyrhizobium japonicum Irr and 2-oxoglutarate binding to Anabaena FurA. How these protein-protein and protein-ligand interactions act in conjunction with regulatory metal ions to facilitate signal integration is an active area of investigation.

Iron homeostasis in Bacillus subtilis relies on three differentially expressed efflux systems.

In Bacillus subtilis, iron homeostasis is maintained by the ferric uptake regulator (Fur) and manganese homeostasis relies on the manganese transport regulator (MntR). Both Fur and MntR function as bi-functional metalloregulators that repress import and activate metal ion efflux systems. The ferrous iron efflux ATPase, PfeT, is derepressed by hydrogen peroxide (H2O2) as sensed by PerR and induced by iron as sensed by Fur. Mutants lacking PfeT are sensitive to iron intoxication. Here, we show that mntR mutants are also iron-sensitive, largely due to decreased expression of the MntR-activated MneP and MneS cation diffusion facilitator (CDF) proteins previously defined for their role in Mn2+ export. The ability of MneP and MneS to export iron is apparent even when their expression is not induced by Mn2+. Our results demonstrate that PfeT, MneP and MneS each contribute to iron homeostasis, and a triple mutant lacking all three is more iron-sensitive than any single mutant. We further show that sensitivity to H2O2 does not correlate with iron sensitivity. For example, an mntR mutant is H2O2-sensitive due to elevated Mn(II) that increases PerR-mediated repression of peroxide resistance genes, and this repression is antagonized by elevated Fe2+ in an mntR pfeT mutant. Thus, H2O2-sensitivity reflects the relative levels of Mn2+ and Fe2+ as sensed by the PerR regulatory protein. These results underscore the complex interplay between manganese, iron and oxidative stress in B. subtilis.

Sulfur availability for Vibrio fischeri growth during symbiosis establishment depends on biogeography within the squid light organ.

The fitness of host-associated microbes depends on their ability to access nutrients in vivo. Identifying these mechanisms is significant for understanding how microbes have evolved to fill specific ecological niches within a host. Vibrio fischeri is a bioluminescent bacterium that colonizes and proliferates within the light organ of the Hawaiian bobtail squid, which provides an opportunity to study how bacteria grow in vivo. Here, the transcription factor CysB is shown to be necessary for V. fischeri both to grow on several sulfur sources in vitro and to establish symbiosis with juvenile squid. CysB is also found to regulate several genes involved in sulfate assimilation and to contribute to the growth of V. fischeri on cystine, which is the oxidized form of cysteine. A mutant that grows on cystine but not sulfate could establish symbiosis, suggesting that V. fischeri acquires nutrients related to this compound within the host. Finally, CysB-regulated genes are shown to be differentially expressed among the V. fischeri populations occupying the various colonization sites found within the light organ. Together, these results suggest the biogeography of V. fischeri populations within the squid light organ impacts the physiology of this symbiotic bacterium in vivo through CysB-dependent gene regulation.

Write a Researcher: a Pen Pal Outreach Program for Middle and High School Students.

The decision to pursue a science, technology, engineering, and/or math (STEM) career is often made in middle and high school, yet many students do not have access to resources or opportunities to navigate this career path. Without guidance, obtaining a job in STEM may seem like a daunting or impossible task. Here, we present Write a Researcher, a program wherein middle and high school students were paired with STEM professionals at Cornell University as pen pals. Through personalized handwritten letters, students her empowered with advice, encouragement, and knowledge to support their understanding of STEM fields, advanced degrees, and careers. We share Write a Researcher here to encourage others to establish similar programs at their own institutions.

Quorum sensing inhibits interference competition among bacterial symbionts within a host.

The symbioses that animals form with bacteria play important roles in health and disease, but the molecular details underlying how bacterial symbionts initially assemble within a host remain unclear.1,2,3 The bioluminescent bacterium Vibrio fischeri establishes a light-emitting symbiosis with the Hawaiian bobtail squid Euprymna scolopes by colonizing specific epithelium-lined crypt spaces within a symbiotic organ called the light organ.4 Competition for these colonization sites occurs between different strains of V. fischeri, with the lancet-like type VI secretion system (T6SS) facilitating strong competitive interference that results in strain incompatibility within a crypt space.5,6 Although recent studies have identified regulators of this T6SS, how the T6SS is controlled as symbionts assemble in vivo remains unknown.7,8 Here, we show that T6SS activity is suppressed by N-octanoyl-L-homoserine lactone (C8 HSL), which is a signaling molecule that facilitates quorum sensing in V. fischeri and is important for efficient symbiont assembly.9,10 We find that this signaling depends on the quorum-sensing regulator LitR, which lowers expression of the needle subunit Hcp, a key component of the T6SS, by repressing transcription of the T6SS regulator VasH. We show that LitR-dependent quorum sensing inhibits strain incompatibility within the squid light organ. Collectively, these results provide new insights into the mechanisms by which regulatory networks that promote symbiosis also control competition among symbionts, which in turn may affect the overall symbiont diversity that assembles within a host.