Increased Intracellular Cyclic di-AMP Levels Sensitize Streptococcus gallolyticus subsp. gallolyticus to Osmotic Stress and Reduce Biofilm Formation and Adherence on Intestinal Cells

Teh, Wooi Keong; Dramsi, Shaynoor; Tolker‐Nielsen, Tim; Yang, Liang; Givskov, Michael

doi:10.1128/jb.00597-18

Cited by 33 publications

(51 citation statements)

References 73 publications

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“…Our success in creating a cdaA -deletion mutant was somewhat unexpected, since it has proven difficult to inactivate the sole diadenylate cyclase CdaA in several streptococci by conventional protocols. Whereas deletion of cdaA was possible in S. pyogenes [ 3 ] and S. mutans [ 45 , 46 ], all attempts to delete cdaA have been unsuccessful in S. pneumoniae [ 15 ] and S. gallolyticus [ 47 ]. In S. agalactiae , it was only possible to delete the chromosomal copy of cdaA in a strain carrying an inducible copy of cdaA on a plasmid [ 44 ].…”

Section: Discussionmentioning

confidence: 99%

Cyclic Di-adenosine Monophosphate Regulates Metabolism and Growth in the Oral Commensal Streptococcus mitis

et al. 2020

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Cyclic di-adenosine monophosphate (c-di-AMP) has emerged as an important bacterial signaling molecule that functions both as an intracellular second messenger in bacterial cells and an extracellular ligand involved in bacteria-host cross-talk. In this study, we identify and characterize proteins involved in controlling the c-di-AMP concentration in the oral commensal and opportunistic pathogen Streptococcusmitis (S. mitis). We identified three known types of c-di-AMP turnover proteins in the genome of S. mitis CCUG31611: a CdaA-type diadenylate cyclase as well as GdpP-, and DhhP-type phosphodiesterases. Biochemical analyses of purified proteins demonstrated that CdaA synthesizes c-di-AMP from ATP whereas both phosphodiesterases can utilize c-di-AMP as well as the intermediary metabolite of c-di-AMP hydrolysis 5′-phosphadenylyl-adenosine (pApA) as substrate to generate AMP, albeit at different catalytic efficiency. Using deletion mutants of each of the genes encoding c-di-AMP turnover proteins, we show by high resolution MS/MS that the intracellular concentration of c-di-AMP is increased in deletion mutants of the phosphodiesterases and non-detectable in the cdaA-mutant. We also detected pApA in mutants of the DhhP-type phosphodiesterase. Low and high levels of c-di-AMP were associated with longer and shorter chains of S. mitis, respectively indicating a role in regulation of cell division. The deletion mutant of the DhhP-type phosphodiesterase displayed slow growth and reduced rate of glucose metabolism.

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Section: Discussionmentioning

confidence: 99%

Cyclic Di-adenosine Monophosphate Regulates Metabolism and Growth in the Oral Commensal Streptococcus mitis

et al. 2020

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“…Previously, we found that inactivation of the main glycine betaine transporter OpuD (SAUSA300_1245) as well as the predicted amino acid transporter AlsT (SAUSA300_1252) allows an S. aureus dacA mutant to grow in rich medium in the absence of c‐di‐AMP (Zeden et al, ). In several other Firmicutes, including B. subtilis, Lactococcus lactis, Streptococcus pneumoniae, S. agalactiae, Streptococcus pyogenes and L. monocytogenes , inactivating mutations have also been identified in osmolyte and potassium transport systems that allow these bacteria to grow in the absence of c‐di‐AMP (Bai et al, ; Corrigan et al, ; Devaux et al, ; Fahmi et al, ; Gundlach, Commichau, et al, ; Gundlach, Herzberg, Hertel, et al, ; Pham et al, ; Pham & Turner, ; Quintana et al, ; Rocha et al, ; Teh et al, ; Whiteley et al, , ; Witte et al, ; Zarrella et al, ; Zeden et al, ) . This suggests that potassium and osmolyte transporters are more active in the absence of c‐di‐AMP, resulting in the accumulation of toxic levels of potassium and osmolytes in the cell.…”

Section: Introductionmentioning

confidence: 99%

Identification of the main glutamine and glutamate transporters in Staphylococcus aureus and their impact on c‐di‐AMP production

Zeden

Kviatkovski

Schuster

et al. 2020

Molecular Microbiology

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A Staphylococcus aureus strain deleted for the c‐di‐AMP cyclase gene dacA is unable to survive in rich medium unless it acquires compensatory mutations. Previously identified mutations were in opuD, encoding the main glycine‐betaine transporter, and alsT, encoding a predicted amino acid transporter. Here, we show that inactivation of OpuD restores the cell size of a dacA mutant to near wild‐type (WT) size, while inactivation of AlsT does not. AlsT was identified as an efficient glutamine transporter, indicating that preventing glutamine uptake in rich medium rescues the growth of the S. aureus dacA mutant. In addition, GltS was identified as a glutamate transporter. By performing growth curves with WT, alsT and gltS mutant strains in defined medium supplemented with ammonium, glutamine or glutamate, we revealed that ammonium and glutamine, but not glutamate promote the growth of S. aureus. This suggests that besides ammonium also glutamine can serve as a nitrogen source under these conditions. Ammonium and uptake of glutamine via AlsT and hence likely a higher intracellular glutamine concentration inhibited c‐di‐AMP production, while glutamate uptake had no effect. These findings provide, besides the previously reported link between potassium and osmolyte uptake, a connection between nitrogen metabolism and c‐di‐AMP signalling in S. aureus.

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“…The present results with pneumococcus grown on solid media (a more biofilm-like condition) contrast with prior studies of these bacteria grown in a strictly planktonic liquid broth culture (26), suggesting that c-di-AMP metabolism varies across types of bacterial growth. This is plausible given the many roles c-di-AMP plays in bacterial physiology, including evidence that c-di-AMP itself helps determine planktonic vs. biofilm organization in streptococci (22,(35)(36)(37)(38). Some virulence factors, like pneumolysin which can induce macrophage cytotoxicity as observed here, are more strongly expressed in biofilm-like growth states of pneumococci (32,33).…”

Section: Discussionmentioning

confidence: 60%

Unique Roles for Streptococcus pneumoniae Phosphodiesterase 2 in Cyclic di-AMP Catabolism and Macrophage Responses

et al. 2020

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Cyclic di-AMP (c-di-AMP) is an important signaling molecule for pneumococci, and as a uniquely prokaryotic product it can be recognized by mammalian cells as a danger signal that triggers innate immunity. Roles of c-di-AMP in directing host responses during pneumococcal infection are only beginning to be defined. We hypothesized that pneumococci with defective c-di-AMP catabolism due to phosphodiesterase deletions could illuminate roles of c-di-AMP in mediating host responses to pneumococcal infection. Pneumococci deficient in phosphodiesterase 2 (Pde2) stimulated a rapid induction of interferon β (IFNβ) expression that was exaggerated in comparison to that induced by wild type (WT) bacteria or bacteria deficient in phosphodiesterase 1. This IFNβ burst was elicited in mouse and human macrophage-like cell lines as well as in primary alveolar macrophages collected from mice with pneumococcal pneumonia. Macrophage hyperactivation by Pde2-deficient pneumococci led to rapid cell death. STING and cGAS were essential for the excessive IFNβ induction, which also required phagocytosis of bacteria and triggered the phosphorylation of IRF3 and IRF7 transcription factors. The select effects of Pde2 deletion were products of a unique role of this enzyme in c-di-AMP catabolism when pneumococci were grown on solid substrate conditions designed to enhance virulence. Because pneumococci with elevated c-di-AMP drive aberrant innate immune responses from macrophages involving hyperactivation of STING, excessive IFNβ expression, and rapid cytotoxicity, we surmise that c-di-AMP is pivotal for directing innate immunity and host-pathogen interactions during pneumococcal pneumonia.

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Increased Intracellular Cyclic di-AMP Levels Sensitize Streptococcus gallolyticus subsp. gallolyticus to Osmotic Stress and Reduce Biofilm Formation and Adherence on Intestinal Cells

Cited by 33 publications

References 73 publications

Cyclic Di-adenosine Monophosphate Regulates Metabolism and Growth in the Oral Commensal Streptococcus mitis

Cyclic Di-adenosine Monophosphate Regulates Metabolism and Growth in the Oral Commensal Streptococcus mitis

Identification of the main glutamine and glutamate transporters in Staphylococcus aureus and their impact on c‐di‐AMP production

Unique Roles for Streptococcus pneumoniae Phosphodiesterase 2 in Cyclic di-AMP Catabolism and Macrophage Responses

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