Cyclic di-AMP Signaling in Bacteria

Stülke, Jörg; Krüger, Larissa

doi:10.1146/annurev-micro-020518-115943

Cited by 128 publications

(167 citation statements)

References 129 publications

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“…Based on a variety of distinct suppressor screens to adapt a B. subtilis strain lacking c-di-AMP to either the presence of glutamate or to growth on complex medium, we can conclude that the control of potassium homeostasis is the major bottleneck that limits growth of the mutant under both conditions. This is in good agreement with (i) the large variety of c-di-AMP target proteins and RNA molecules that are involved in the uptake and export of potassium in B. subtilis and other bacteria [11,13,20,21,23,31,34,45,78], (ii) the fact that the intracellular c-di-AMP levels seem to report the extracellular potassium concentrations [18,21,37], and the isolation of suppressor mutants affecting potassium homeostasis in response to altered cellular c-di-AMP levels also in other bacteria [18,37]. However, our study also clearly demonstrates that potassium is not the only problem for growth of B. subtilis in the absence of c-di-AMP since the presence of mutations that reduce potassium uptake or facilitate its export is not sufficient to overcome the toxicity of glutamate or complex medium.…”

Section: Aima (Ybec) Is a Novel Major Glutamate Transportersupporting

confidence: 81%

Essentiality of c-di-AMP inBacillus subtilis: Bypassing mutations converge in potassium and glutamate homeostasis

Krüger

Herzberg

Rath

et al. 2020

Preprint

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In order to adjust to changing environmental conditions, bacteria use nucleotide second messengers to transduce external signals and translate them into a specific cellular response. Cyclic di-adenosine monophosphate (c-di-AMP) is the only known essential nucleotide second messenger. In addition to the well-established role of this second messenger in the control of potassium homeostasis, we observed that glutamate is as toxic as potassium for a c-di-AMP-free strain of the Gram-positive model bacterium Bacillus subtilis . In this work, we isolated suppressor mutants that allow growth of a c-di-AMP-free strain under these toxic conditions. Characterization of glutamate resistant suppressors revealed that they contain pairs of mutations, in most cases affecting glutamate and potassium homeostasis. Among these mutations, several independent mutations affected a novel glutamate transporter, AimA ( A mino acid im porter A , formerly YbeC). This protein is the major transporter for glutamate and serine in B. subtilis . Unexpectedly, some of the isolated suppressor mutants could suppress glutamate toxicity by a combination of mutations that affect phospholipid biosynthesis and a specific gain-of-function mutation of a mechanosensitive channel of small conductance (YfkC) suggesting the acquisition of a device for glutamate export. Cultivation of the c-di-AMP-free strain on complex medium was an even greater challenge because the amounts of potassium, glutamate, and other osmolytes are substantially higher than in minimal medium. Suppressor mutants viable on complex medium could only be isolated under anaerobic conditions if one of the two c-di-AMP receptor proteins, DarA or DarB, was absent. Also on complex medium, potassium and osmolyte toxicity are the major bottlenecks for the growth of B. subtilis in the absence of c-di-AMP. Our results indicate that the essentiality of c-di-AMP in B. subtilis is caused by the global impact of the second messenger nucleotide on different aspects of cellular physiology.

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Section: Aima (Ybec) Is a Novel Major Glutamate Transportersupporting

confidence: 81%

Essentiality of c-di-AMP inBacillus subtilis: Bypassing mutations converge in potassium and glutamate homeostasis

Krüger

Herzberg

Rath

et al. 2020

Preprint

Self Cite

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“…For many moonlighting enzymes, the signals controlling the secondary function of the enzymes have been identified (Commichau and Stülke, 2008). However, this is not the case for GlmM, which controls the activity of CdaA and thus the uptake of osmolytes via transporters whose activities are regulated by c-di-AMP (Stülke and Krüger, 2020).…”

Section: Discussionmentioning

confidence: 99%

“…So how does a cell synthesizing CdaA and GlmM sense osmotic up-and downshifts and how is c-di-AMP production regulated by employing the phosphoglucosamine mutase? Over the past years, it has been observed that in many bacteria cdi-AMP is controlling the uptake and efflux of potassium ions to adjust the cellular turgor (Stülke and Krüger, 2020). Thus, the cellular potassium concentration could control the regulatory interaction between GlmM and CdaA.…”

Section: Discussionmentioning

confidence: 99%

“…By contrast, a low intracellular potassium concentration might trigger the inhibition of CdaA by GlmM to allow the uptake of the osmolyte and the adjustment of the cellular turgor. However, in addition to potassium, other osmolytes like glycine betaine and carnitine are taken up by c-di-AMPregulated transporters (Commichau et al, 2018;Stülke and Krüger, 2020). Therefore, it can be assumed that a mechanism, which is independent of a specific osmolyte, ensures the adjustment of the cellular turgor.…”

Section: Discussionmentioning

confidence: 99%

“…Cyclic di-AMP (c-di-AMP) plays a central role in the adaptation of bacteria to the environmental osmolarity (Witte et al, 2008;Commichau et al, 2015Commichau et al, , 2018Commichau et al, , 2019Fahmi et al, 2017;Stülke and Krüger, 2020). c-di-AMP is in fact essential for Bacillus subtilis, Lactococcus lactis, Listeria monocytogenes and other Gram-positive bacteria (Woodward et al, 2010;Corrigan et al, 2011;Luo and Helmann, 2012;Bai et al, 2013;Mehne et al, 2013;Witte et al, 2013;Whiteley et al, 2015;Rismondo et al, 2016) since the dinucleotide prevents the uptake of potassium and other osmolytes to toxic levels by direct binding to the respective transport systems Bai et al, 2014;Kim et al, 2015;Schuster et al, 2016;Blötz et al, 2017;Gundlach et al, 2017;Whiteley et al, 2017;Devaux et al, 2018;Pham et al, 2018;Zarella et al, 2018;Zeden et al, 2018;Gibhardt et al, 2019;Gundlach et al, 2019;Quintana et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

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An extracytoplasmic protein and a moonlighting enzyme modulate synthesis of c‐di‐AMP in Listeria monocytogenes

Gibhardt

Heidemann

Bremenkamp

et al. 2020

Environmental Microbiology

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Summary The second messenger cyclic di‐AMP (c‐di‐AMP) is essential for growth of many bacteria because it controls osmolyte homeostasis. c‐di‐AMP can regulate the synthesis of potassium uptake systems in some bacteria and it also directly inhibits and activates potassium import and export systems, respectively. Therefore, c‐di‐AMP production and degradation have to be tightly regulated depending on the environmental osmolarity. The Gram‐positive pathogen Listeria monocytogenes relies on the membrane‐bound diadenylate cyclase CdaA for c‐di‐AMP production and degrades the nucleotide with two phosphodiesterases. While the enzymes producing and degrading the dinucleotide have been reasonably well examined, the regulation of c‐di‐AMP production is not well understood yet. Here we demonstrate that the extracytoplasmic regulator CdaR interacts with CdaA via its transmembrane helix to modulate c‐di‐AMP production. Moreover, we show that the phosphoglucosamine mutase GlmM forms a complex with CdaA and inhibits the diadenylate cyclase activity in vitro. We also found that GlmM inhibits c‐di‐AMP production in L. monocytogenes when the bacteria encounter osmotic stress. Thus, GlmM is the major factor controlling the activity of CdaA in vivo. GlmM can be assigned to the class of moonlighting proteins because it is active in metabolism and adjusts the cellular turgor depending on environmental osmolarity.

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A Luminescence‐Based Coupled Enzyme Assay Enables High‐Throughput Quantification of the Bacterial Second Messenger 3’3’‐Cyclic‐Di‐AMP

Zaver

Pollock

Boradia³

et al. 2020

ChemBioChem

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Cyclic dinucleotide signaling systems, which are found ubiquitously throughout nature, allow organisms to rapidly and dynamically sense and respond to alterations in their environments. In recent years, the second messenger, cyclic di‐(3’,5’)‐adenosine monophosphate (c‐di‐AMP), has been identified as an essential signaling molecule in a diverse array of bacterial genera. We and others have shown that defects in c‐di‐AMP homeostasis result in severe physiological defects and virulence attenuation in many bacterial species. Despite significant advancements in the field, there is still a major gap in the understanding of the environmental and cellular factors that influence c‐di‐AMP dynamics due to a lack of tools to sensitively and rapidly monitor changes in c‐di‐AMP levels. To address this limitation, we describe here the development of a luciferase‐based coupled enzyme assay that leverages the cyclic nucleotide phosphodiesterase, CnpB, for the sensitive and high‐throughput quantification of 3’3’‐c‐di‐AMP. We also demonstrate the utility of this approach for the quantification of the cyclic oligonucleotide‐based anti‐phage signaling system (CBASS) effector, 3’3’‐cGAMP. These findings establish CDA‐Luc as a more affordable and sensitive alternative to conventional c‐di‐AMP detection tools with broad utility for the study of bacterial cyclic dinucleotide physiology.

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Cyclic di-AMP Signaling in Bacteria

Cited by 128 publications

References 129 publications

Essentiality of c-di-AMP inBacillus subtilis: Bypassing mutations converge in potassium and glutamate homeostasis

Essentiality of c-di-AMP inBacillus subtilis: Bypassing mutations converge in potassium and glutamate homeostasis

An extracytoplasmic protein and a moonlighting enzyme modulate synthesis of c‐di‐AMP in Listeria monocytogenes

A Luminescence‐Based Coupled Enzyme Assay Enables High‐Throughput Quantification of the Bacterial Second Messenger 3’3’‐Cyclic‐Di‐AMP

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