Listeria monocytogenes Multidrug Resistance Transporters and Cyclic Di-AMP, Which Contribute to Type I Interferon Induction, Play a Role in Cell Wall Stress

Zeevi, Millie Kaplan; Shafir, Nirit S.; Shaham, Shira; Friedman, Sivan; Sigal, Nadejda; Nir-Paz, Ran; Boneca, Ivo Gomperts; Herskovits, Anat A.

doi:10.1128/jb.00794-13

Cited by 60 publications

(66 citation statements)

References 56 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To directly evaluate the interaction between the domains of PgpH and c-di-AMP, we used a differential radial action of ligand assay (DRACALA), which tests for protein binding with radiolabeled nucleotides (25). These binding experiments indicated that the ED domain and MBP alone did not interact with 32 P-c-di-AMP, whereas the MBP-HD domain construct exhibited clear binding (Fig. 1D).…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

An HD-domain phosphodiesterase mediates cooperative hydrolysis of c-di-AMP to affect bacterial growth and virulence

Huynh

Luo

Pensinger

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

174

252

View full text Add to dashboard Cite

The nucleotide cyclic di-3′,5′- adenosine monophosphate (c-di-AMP) was recently identified as an essential and widespread second messenger in bacterial signaling. Among c-di-AMP–producing bacteria, altered nucleotide levels result in several physiological defects and attenuated virulence. Thus, a detailed molecular understanding of c-di-AMP metabolism is of both fundamental and practical interest. Currently, c-di-AMP degradation is recognized solely among DHH-DHHA1 domain-containing phosphodiesterases. Using chemical proteomics, we identified the Listeria monocytogenes protein PgpH as a molecular target of c-di-AMP. Biochemical and structural studies revealed that the PgpH His-Asp (HD) domain bound c-di-AMP with high affinity and specifically hydrolyzed this nucleotide to 5′-pApA. PgpH hydrolysis activity was inhibited by ppGpp, indicating a cross-talk between c-di-AMP signaling and the stringent response. Genetic analyses supported coordinated regulation of c-di-AMP levels in and out of the host. Intriguingly, a L. monocytogenes mutant that lacks c-di-AMP phosphodiesterases exhibited elevated c-di-AMP levels, hyperinduced a host type-I IFN response, and was significantly attenuated for infection. Furthermore, PgpH homologs, which belong to the 7TMR-HD family, are widespread among hundreds of c-di-AMP synthesizing microorganisms. Thus, PgpH represents a broadly conserved class of c-di-AMP phosphodiesterase with possibly other physiological functions in this crucial signaling network.

show abstract

Section: Resultsmentioning

confidence: 99%

“…To evaluate binding specificity, we conducted competition assays, in which unlabeled nucleotides were added in excess to disrupt 32 P-c-di-AMP interaction with MBP-HD protein. Among the nucleotides tested, only 5′-pApA and c-di-GMP diminished radiolabeled c-di-AMP binding, although much less than unlabeled c-di-AMP (Fig.…”

Section: Resultsmentioning

confidence: 99%

An HD-domain phosphodiesterase mediates cooperative hydrolysis of c-di-AMP to affect bacterial growth and virulence

Huynh

Luo

Pensinger

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

174

252

View full text Add to dashboard Cite

show abstract

“…While c-di-AMP has been found to regulate many important functions in bacteria, including cell wall stress and peptidoglycan homeostasis (11)(12)(13)(14)(15), antibiotic resistance (16)(17)(18), biofilm formation (19), growth and metabolism (20)(21)(22)(23), and DNA damage response (24,25), it can also modulate leukocyte activation. For example, c-di-AMP secretion into the macrophage cytosol induces a robust type I interferon (IFN [IFN-␤]) response during intracellular bacterial infections, including those caused by Listeria monocytogenes (26,27), Chlamydia trachomatis (28), and Mycobacterium spp.…”

mentioning

confidence: 99%

Cyclic di-AMP Released from Staphylococcus aureus Biofilm Induces a Macrophage Type I Interferon Response

et al. 2016

View full text Add to dashboard Cite

Staphylococcus aureus is a leading cause of community-and nosocomial-acquired infections, with a propensity for biofilm formation. S. aureus biofilms actively skew the host immune response toward an anti-inflammatory state; however, the biofilm effector molecules and the mechanism(s) of action responsible for this phenomenon remain to be fully defined. The essential bacterial second messenger cyclic diadenylate monophosphate (c-di-AMP) is an emerging pathogen-associated molecular pattern during intracellular bacterial infections, as c-di-AMP secretion into the infected host cytosol induces a robust type I interferon (IFN) response. Type I IFNs have the potential to exacerbate infectious outcomes by promoting anti-inflammatory effects; however, the type I IFN response to S. aureus biofilms is unknown. Additionally, while several intracellular proteins function as c-di-AMP receptors in S. aureus, it has yet to be determined if any extracellular role for c-di-AMP exists and its release during biofilm formation has not yet been demonstrated. This study examined the possibility that c-di-AMP released during S. aureus biofilm growth polarizes macrophages toward an anti-inflammatory phenotype via type I interferon signaling. DacA, the enzyme responsible for c-di-AMP synthesis in S. aureus, was highly expressed during biofilm growth, and 30 to 50% of total c-di-AMP produced from S. aureus biofilm was released extracellularly due to autolytic activity. S. aureus biofilm c-di-AMP release induced macrophage type I IFN expression via a STING-dependent pathway and promoted S. aureus intracellular survival in macrophages. These findings identify c-di-AMP as another mechanism for how S. aureus biofilms promote macrophage anti-inflammatory activity, which likely contributes to biofilm persistence. Staphylococcus aureus is a leading cause of prosthetic joint infections (1, 2), whereupon adherence to the implant surface facilitates biofilm formation. These infections are particularly challenging to treat and typically require a two-stage surgical process for insertion of a new device (1). Moreover, biofilms actively skew the host immune response toward an anti-inflammatory state, thereby contributing to the chronic nature of biofilm-mediated infections (3-5). This is evident by macrophage polarization toward an alternatively activated phenotype and the recruitment of myeloid-derived suppressor cells (MDSCs) (3,(6)(7)(8). While this immune deviation appears to be driven by S. aureus biofilm products, the effectors and their mechanism(s) of action remain to be fully identified.To further understand mechanisms of immune modification by S. aureus biofilms, we investigated the role of the essential bacterial second messenger cyclic diadenylate monophosphate (c-di-AMP), since it has been identified as an emerging pathogen-associated molecular pattern (PAMP) that influences immune responsiveness (9, 10). While c-di-AMP has been found to regulate many important functions in bacteria, including cell wall stress and peptidoglycan homeostas...

show abstract

“…Further studies have identified cyclic di-AMP (c-di-AMP) as a substrate of MdrM and as the ligand that triggers the innate immune system to express type I interferons (46,50). While immune cells rapidly sense this cyclic dinucleotide as a signal for bacterial invasion, within the bacteria it was shown to play a regulatory role in cell wall stress responses and homeostasis (49,51).…”

mentioning

confidence: 99%

“…We have previously shown that activation of the type I interferon response by L. monocytogenes relies on the expression of a set of bacterial multidrug-resistant transporters-MdrM, MdrT, MdrA, and MdrC-that together are responsible for most of the response in murine macrophages (48,49). Among these transporters, MdrM was found to be most critical, as deletion of its gene alone led to ϳ70% reduction in IFN-␤ induction compared to that induced by wild-type bacteria.…”

mentioning

confidence: 99%

The Human P-Glycoprotein Transporter Enhances the Type I Interferon Response to Listeria monocytogenes Infection

et al. 2015

View full text Add to dashboard Cite

bHuman multidrug efflux transporters are known for their ability to extrude antibiotics and toxic compounds out of cells, yet accumulating data indicate they have additional functions in diverse physiological processes not related to drug efflux. Here, we show that the human multidrug transporter P-glycoprotein (P-gp) (also named MDR1 and ABCB1) is transcriptionally induced in the monocytic cell line THP-1 upon infection with the human intracellular bacterial pathogen Listeria monocytogenes. Notably, we found that P-gp is important for full activation of the type I interferon response elicited against L. monocytogenes bacteria. Both inhibition of P-gp function by verapamil and inhibition of its transcription using mRNA silencing led to a reduction in the magnitude of the type I response in infected cells. This function of P-gp was specific to type I interferon cytokines elicited against cytosolic replicating bacteria and was not observed in response to cyclic di-AMP (c-di-AMP), a molecule that was shown to be secreted by L. monocytogenes during infection and to trigger type I interferons. Moreover, P-gp was not involved in activation of other proinflammatory cytokines, such as those triggered by vacuolar-restricted L. monocytogenes or lipopolysaccharide (LPS). Taken together, these findings demonstrate a role for P-gp in proper development of an innate immune response against intracellular pathogens, highlighting the complexity in employing therapeutic strategies that involve inhibition of multidrug resistance (MDR) efflux pumps. Multidrug transporters mediate the active efflux of a wide range of drugs and xenobiotics, including antibiotics and chemotherapeutics (1). This permissive efflux ability engenders multidrug resistance (MDR)-a phenomenon that largely underlies the failure of various chemotherapeutic treatments (2-4). Human MDR transporters harbor an ATP-binding cassette (ABC), which defines the ABC-type superfamily, comprising more than 45 proteins in the human genome (5). Among these, several transporters have been extensively studied, such as the P-glycoprotein (P-gp) (also named MDR1 and ABCB1) (6), BCRP (ABCG2) (7), and MRP1 (ABCC1) (8), which were all shown to exhibit clinically relevant MDR functions (9). P-gp, encoded by the MDR1 gene, is the most prominent and best-characterized member of the ABCtype superfamily, first isolated in clinical cancers (6, 10). Aside from its well-documented multidrug resistance function in cancer cells, P-gp is naturally expressed in a variety of normal tissues and cells, including immune cells, such as macrophages, dendritic cells, T and B lymphocytes, and natural killer (NK) cells, and was shown to possess physiological functions beyond detoxification (11-15). Several studies have indicated roles for P-gp in lipid transport, intracellular trafficking of cholesterol, cell death, cell differentiation, and immune responses (16,17). Regarding the last, P-gp was shown to exhibit immunomodulatory activity and to influence the secretion of various inflammatory mediators,...

show abstract

Listeria monocytogenes Multidrug Resistance Transporters and Cyclic Di-AMP, Which Contribute to Type I Interferon Induction, Play a Role in Cell Wall Stress

Cited by 60 publications

References 56 publications

An HD-domain phosphodiesterase mediates cooperative hydrolysis of c-di-AMP to affect bacterial growth and virulence

An HD-domain phosphodiesterase mediates cooperative hydrolysis of c-di-AMP to affect bacterial growth and virulence

Cyclic di-AMP Released from Staphylococcus aureus Biofilm Induces a Macrophage Type I Interferon Response

The Human P-Glycoprotein Transporter Enhances the Type I Interferon Response to Listeria monocytogenes Infection

Contact Info

Product

Resources

About