Cyclic di-GMP-dependent Signaling Pathways in the Pathogenic Firmicute Listeria monocytogenes

Köseoğlu, Volkan K.; Güvener, Zehra Tüzün; Myers-Morales, Tanya; Reed, Joseph M.; D’Orazio, Sarah E. F.; Miller, Kurt W.; Gomelsky, Mark

doi:10.1371/journal.ppat.1004301

Cited by 84 publications

(129 citation statements)

References 96 publications

Supporting

Mentioning

120

Contrasting

Order By: Relevance

“…This finding indicates that inhibition of motility can be controlled by FlgZ and highlights a role for Pel polysaccharide in this process. The finding that Pel polysaccharide plays a role in motility repression in the absence of c-di-GMP effector protein FlgZ, particularly in the ⌬bifA ⌬flgZ and ⌬hptB ⌬flgZ mutants, aligns with previous findings indicating that extracellular polysaccharides impair motility in other species, including S. enterica (19), B. subtilis (43), and L. monocytogenes (44). However, P. aeruginosa is different from these and other organisms using c-di-GMP-dependent motility control in that eliminating either the c-di-GMP binding protein or the exopolysaccharide singly has no significant impact on motility in the high-c-di-GMP background.…”

Section: Discussionsupporting

confidence: 90%

“…Pel plays important roles in cell aggregation, and we hypothesized that elevated levels of Pel may be inhibiting cell mobility and masking any contributions of FlgZ to regulating flagellum-mediated swarming motility in the ⌬bifA mutant. Similarly, other extracellular polysaccharides have been shown to impair cell motility in S. enterica (19), B. subtilis (43), and Listeria monocytogenes (44).…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

PilZ Domain Protein FlgZ Mediates Cyclic Di-GMP-Dependent Swarming Motility Control in Pseudomonas aeruginosa

et al. 2016

View full text Add to dashboard Cite

The second messenger cyclic diguanylate (c-di-GMP) is an important regulator of motility in many bacterial species. In Pseudomonas aeruginosa, elevated levels of c-di-GMP promote biofilm formation and repress flagellum-driven swarming motility. The rotation of P. aeruginosa's polar flagellum is controlled by two distinct stator complexes, MotAB, which cannot support swarming motility, and MotCD, which promotes swarming motility. Here we show that when c-di-GMP levels are elevated, swarming motility is repressed by the PilZ domain-containing protein FlgZ and by Pel polysaccharide production. We demonstrate that FlgZ interacts specifically with the motility-promoting stator protein MotC in a c-di-GMP-dependent manner and that a functional green fluorescent protein (GFP)-FlgZ fusion protein shows significantly reduced polar localization in a strain lacking the MotCD stator. Our results establish FlgZ as a c-di-GMP receptor affecting swarming motility by P. aeruginosa and support a model wherein c-di-GMP-bound FlgZ impedes motility via its interaction with the MotCD stator. IMPORTANCEThe regulation of surface-associated motility plays an important role in bacterial surface colonization and biofilm formation. c-di-GMP signaling is a widespread means of controlling bacterial motility, and yet the mechanism whereby this signal controls surface-associated motility in P. aeruginosa remains poorly understood. Here we identify a PilZ domain-containing c-di-GMP effector protein that contributes to c-di-GMP-mediated repression of swarming motility by P. aeruginosa. We provide evidence that this effector, FlgZ, impacts swarming motility via its interactions with flagellar stator protein MotC. Thus, we propose a new mechanism for c-di-GMP-mediated regulation of motility for a bacterium with two flagellar stator sets, increasing our understanding of surface-associated behaviors, a key prerequisite to identifying ways to control the formation of biofilm communities. Cyclic diguanylate (c-di-GMP) is a ubiquitous bacterial second messenger responsible for regulating a range of cellular processes, including motility and biofilm formation (1). In general, low intracellular c-di-GMP levels are associated with motile lifestyles, while elevated levels of c-di-GMP promote surface attachment and sessile lifestyles (2, 3). c-di-GMP is synthesized from two molecules of GTP by diguanylate cyclases (DGCs) and degraded by c-di-GMP-specific phosphodiesterases (PDEs) (1, 4). Many DGCs and PDEs involved in motility regulation have been characterized, but the mechanisms by which c-di-GMP regulates motility are poorly understood in Pseudomonas aeruginosa and in other bacterial species.To regulate numerous biological functions, c-di-GMP binds to specific effector proteins or RNA (reviewed in reference 5). Recent studies have focused on identifying these c-di-GMP effectors and their mechanisms for regulating c-di-GMP-dependent processes. One class of effectors is the PilZ domain-containing protein family, which is characterized by conserved ...

show abstract

Section: Discussionsupporting

confidence: 90%

Section: Resultsmentioning

confidence: 99%

PilZ Domain Protein FlgZ Mediates Cyclic Di-GMP-Dependent Swarming Motility Control in Pseudomonas aeruginosa

et al. 2016

View full text Add to dashboard Cite

show abstract

“…Similar effects of YhjH orthologs have been reported in Aeromonas veronii, Dickeya dadantii, and Klebsiella pneumoniae (27)(28)(29). More recently, three EAL-only proteins from the Gram-positive pathogen Listeria monocytogenes were shown to possess c-di-GMP PDE activity (30).…”

supporting

confidence: 63%

Stand-Alone EAL Domain Proteins Form a Distinct Subclass of EAL Proteins Involved in Regulation of Cell Motility and Biofilm Formation in Enterobacteria

et al. 2017

View full text Add to dashboard Cite

The second messenger cyclic dimeric GMP (c-di-GMP) is almost ubiquitous among bacteria as are the c-di-GMP turnover proteins, which mediate the transition between motility and sessility. EAL domain proteins have been characterized as c-di-GMP-specific phosphodiesterases. While most EAL domain proteins contain additional, usually N-terminal, domains, there is a distinct family of proteins with stand-alone EAL domains, exemplified by Salmonella enterica serovar Typhimurium proteins STM3611 (YhjH/PdeH), a c-di-GMP-specific phosphodiesterase, and the enzymatically inactive STM1344 (YdiV/CdgR) and STM1697, which regulate bacterial motility through interaction with the flagellar master regulator, FlhDC. We have analyzed the phylogenetic distribution of EAL-only proteins and their potential functions. Genes encoding EAL-only proteins were found in various bacterial phyla, although most of them were seen in proteobacteria, particularly enterobacteria. Based on the conservation of the active site residues, nearly all stand-alone EAL domains encoded by genomes from phyla other than proteobacteria appear to represent functional phosphodiesterases. Within enterobacteria, EAL-only proteins were found to cluster either with YhjH or with one of the subfamilies of YdiV-related proteins. EAL-only proteins from Shigella flexneri, Klebsiella pneumoniae, and Yersinia enterocolitica were tested for their ability to regulate swimming and swarming motility and formation of the red, dry, and rough (rdar) biofilm morphotype. In these tests, YhjH-related proteins S4210, KPN_01159, KPN_03274, and YE4063 displayed properties typical of enzymatically active phosphodiesterases, whereas S1641 and YE1324 behaved like members of the YdiV/STM1697 subfamily, with Yersinia enterocolitica protein YE1324 shown to downregulate motility in its native host. Of two closely related EAL-only proteins, YE2225 is an active phosphodiesterase, while YE1324 appears to interact with FlhD. These results suggest that in FlhDC-harboring beta-and gammaproteobacteria, some EAL-only proteins evolved to become catalytically inactive and regulate motility and biofilm formation by interacting with FlhDC.IMPORTANCE The EAL domain superfamily consists mainly of proteins with cyclic dimeric GMP-specific phosphodiesterase activity, but individual domains have been classified in three classes according to their functions and conserved amino acid signatures. Proteins that consist solely of stand-alone EAL domains cannot rely on other domains to form catalytically active dimers, and most of them fall into one of two distinct classes: catalytically active phosphodiesterases with well-conserved residues of the active site and the dimerization loop, and catalytically inactive YdiV/ CdgR-like proteins that regulate bacterial motility by binding to the flagellar master

show abstract

“…The expression in this reporter strain of heterologous PDEs from diverse bacteria can partially rescue swimming in semisolid agar (37)(38)(39). The MBP fusions to both BldP1 from M. marinus (Fig.…”

Section: Resultsmentioning

confidence: 98%

Optogenetic Module for Dichromatic Control of c-di-GMP Signaling

et al. 2017

Self Cite

View full text Add to dashboard Cite

Many aspects of bacterial physiology and behavior, including motility, surface attachment, and the cell cycle, are controlled by cyclic di-GMP (c-di-GMP)-dependent signaling pathways on the scale of seconds to minutes. Interrogation of such processes in real time requires tools for introducing rapid and reversible changes in intracellular c-di-GMP levels. Inducing the expression of genes encoding c-di-GMP-synthetic (diguanylate cyclases) and -degrading (c-di-GMP phosphodiesterase) enzymes by chemicals may not provide adequate temporal control. In contrast, light-controlled diguanylate cyclases and phosphodiesterases can be quickly activated and inactivated. A red/near-infrared-light-regulated diguanylate cyclase, BphS, was engineered previously, yet a complementary light-activated c-di-GMP phosphodiesterase has been lacking. In search of such a phosphodiesterase, we investigated two homologous proteins from Allochromatium vinosum and Magnetococcus marinus, designated BldP, which contain C-terminal EAL-BLUF modules, where EAL is a c-di-GMP phosphodiesterase domain and BLUF is a blue light sensory domain. Characterization of the BldP proteins in Escherichia coli and in vitro showed that they possess light-activated c-di-GMP phosphodiesterase activities. Interestingly, light activation in both enzymes was dependent on oxygen levels. The truncated EAL-BLUF fragment from A. vinosum BldP lacked phosphodiesterase activity, whereas a similar fragment from M. marinus BldP, designated EB1, possessed such activity that was highly (Ͼ30-fold) upregulated by light. Following light withdrawal, EB1 reverted to the inactive ground state with a half-life of ϳ6 min. Therefore, the bluelight-activated phosphodiesterase EB1 can be used in combination with the red/nearinfrared-light-regulated diguanylate cyclase BphS for the bidirectional regulation of c-di-GMP-dependent processes in E. coli as well as other bacterial and nonbacterial cells.IMPORTANCE Regulation of motility, attachment to surfaces, the cell cycle, and other bacterial processes controlled by the c-di-GMP signaling pathways occur at a fast (seconds-to-minutes) pace. Interrogation of these processes at high temporal and spatial resolution using chemicals is difficult or impossible, while optogenetic approaches may prove useful. We identified and characterized a robust, blue-lightactivated c-di-GMP phosphodiesterase (hydrolase) that complements a previously engineered red/near-infrared-light-regulated diguanylate cyclase (c-di-GMP synthase). These two enzymes form a dichromatic module for manipulating intracellular c-di-GMP levels in bacterial and nonbacterial cells.KEYWORDS biofilms, c-di-GMP, diguanylate cyclase, gene expression, motility, optogenetics, phosphodiesterase, photoreceptors, photosensory reception, signal transduction S ignaling via second messengers occurs at various time scales, from seconds to hours. To control relatively slow processes, intracellular concentrations of a second messenger can be modulated by the constitutive or inducible (over)e...

show abstract

Cyclic di-GMP-dependent Signaling Pathways in the Pathogenic Firmicute Listeria monocytogenes

Cited by 84 publications

References 96 publications

PilZ Domain Protein FlgZ Mediates Cyclic Di-GMP-Dependent Swarming Motility Control in Pseudomonas aeruginosa

PilZ Domain Protein FlgZ Mediates Cyclic Di-GMP-Dependent Swarming Motility Control in Pseudomonas aeruginosa

Stand-Alone EAL Domain Proteins Form a Distinct Subclass of EAL Proteins Involved in Regulation of Cell Motility and Biofilm Formation in Enterobacteria

Optogenetic Module for Dichromatic Control of c-di-GMP Signaling

Contact Info

Product

Resources

About