Cyclic di-AMP Signaling in Streptococcus pneumoniae

  • Tiffany M. Zarrella
  • Guangchun BaiEmail author


Streptococcus pneumoniae causes diseases such as pneumonia, otitis media, meningitis, and bacteremia. As such, this pathogen survives and adapts to different environmental stimuli and withstands stress conditions encountered during colonization, dissemination, and infection in the respective host compartments. Recent studies designate the bacterial signaling nucleotide cyclic di-adenosine monophosphate (cyclic di-AMP) as an important facet to pneumococcal physiology and virulence. In this chapter, we will describe the signaling network and the role of cyclic di-AMP as a second messenger in pneumococci. In S. pneumoniae, cyclic di-AMP is produced by a sole diadenylate cyclase, CdaA, and is catabolized by two phosphodiesterases, Pde1 and Pde2. cyclic di-AMP is secreted through an unidentified mechanism which may impact host–pathogen interactions. The gene encoding CdaA is essential, and perturbation of cyclic di-AMP levels affects adaptation to stress, epithelial cell adhesion, and pneumococcal virulence, demonstrating that cyclic di-AMP is a pervasive molecule in pathogenesis. A Trk-family cyclic di-AMP binding protein, CabP, has been characterized as a mediator of potassium uptake via the transporter TrkH. Potassium levels affect expression of CdaA, and CabP modulates cyclic di-AMP homeostasis, suggesting that cyclic di-AMP plays a fundamental role in ion transport. Nevertheless, repercussions of cyclic di-AMP signaling discussed here allude to the existence of additional cyclic di-AMP effectors. Future avenues of research and outlying questions of interest are addressed.


Streptococcus pneumoniae Cyclic di-AMP Stress response CdaA Pde1 Pde2 CabP TrkH 



The writing of this chapter by TZ was supported in part by the Intramural Research Program of the National Institutes of Health (NIH), National Cancer Institute, Center for Cancer Research. GB is a subrecipient of NIH grant R35HL135756.

Conflict of Interest

The authors declare no conflict of interest.


  1. 1.
    CDC (2015) Pneumococcal disease. In: Hamborsky J, Kroger A, Wolfe S (eds) Epidemiology and prevention of vaccine-preventable diseases, 13th edn. Public Health Foundation, Washington, DC, pp 279–296Google Scholar
  2. 2.
    Steel HC, Cockeran R, Anderson R, Feldman C (2013) Overview of community-acquired pneumonia and the role of inflammatory mechanisms in the immunopathogenesis of severe pneumococcal disease. Mediat Inflamm 2013:490346. CrossRefGoogle Scholar
  3. 3.
    Gomez-Mejia A, Gamez G, Hammerschmidt S (2017) Streptococcus pneumoniae two-component regulatory systems: the interplay of the pneumococcus with its environment. Int J Med Microbiol. CrossRefGoogle Scholar
  4. 4.
    Yesilkaya H, Andisi VF, Andrew PW, Bijlsma JJ (2013) Streptococcus pneumoniae and reactive oxygen species: an unusual approach to living with radicals. Trends Microbiol 21(4):187–195. CrossRefPubMedGoogle Scholar
  5. 5.
    Kwon HY, Kim SW, Choi MH, Ogunniyi AD, Paton JC, Park SH, Pyo SN, Rhee DK (2003) Effect of heat shock and mutations in ClpL and ClpP on virulence gene expression in Streptococcus pneumoniae. Infect Immun 71(7):3757–3765CrossRefGoogle Scholar
  6. 6.
    Hajaj B, Yesilkaya H, Benisty R, David M, Andrew PW, Porat N (2012) Thiol peroxidase is an important component of Streptococcus pneumoniae in oxygenated environments. Infect Immun 80(12):4333–4343. CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Spellerberg B, Cundell DR, Sandros J, Pearce BJ, Idanpaan-Heikkila I, Rosenow C, Masure HR (1996) Pyruvate oxidase, as a determinant of virulence in Streptococcus pneumoniae. Mol Microbiol 19(4):803–813CrossRefGoogle Scholar
  8. 8.
    Rai P, Parrish M, Tay IJ, Li N, Ackerman S, He F, Kwang J, Chow VT, Engelward BP (2015) Streptococcus pneumoniae secretes hydrogen peroxide leading to DNA damage and apoptosis in lung cells. Proc Natl Acad Sci USA 112(26):E3421–E3430. CrossRefPubMedGoogle Scholar
  9. 9.
    Bassoe CF, Bjerknes R (1985) Phagocytosis by human leukocytes, phagosomal pH and degradation of seven species of bacteria measured by flow cytometry. J Med Microbiol 19(1):115–125. CrossRefPubMedGoogle Scholar
  10. 10.
    Andersen NE, Gyring J, Hansen AJ, Laursen H, Siesjo BK (1989) Brain acidosis in experimental pneumococcal meningitis. J Cereb Blood Flow Metab 9(3):381–387. CrossRefPubMedGoogle Scholar
  11. 11.
    Pilishvili T, Lexau C, Farley MM, Hadler J, Harrison LH, Bennett NM, Reingold A, Thomas A, Schaffner W, Craig AS, Smith PJ, Beall BW, Whitney CG, Moore MR, Active Bacterial Core Surveillance/Emerging Infections Program N (2010) Sustained reductions in invasive pneumococcal disease in the era of conjugate vaccine. J Infect Dis 201(1):32–41. CrossRefPubMedGoogle Scholar
  12. 12.
    Henrichsen J (1995) Six newly recognized types of Streptococcus pneumoniae. J Clin Microbiol 33(10):2759–2762CrossRefGoogle Scholar
  13. 13.
    Yother J (2011) Capsules of Streptococcus pneumoniae and other bacteria: paradigms for polysaccharide biosynthesis and regulation. Annu Rev Microbiol 65:563–581. CrossRefPubMedGoogle Scholar
  14. 14.
    Ortqvist A (2001) Pneumococcal vaccination: current and future issues. Eur Respir J 18(1):184–195CrossRefGoogle Scholar
  15. 15.
    Obolski U, Lourenco J, Thompson C, Thompson R, Gori A, Gupta S (2018) Vaccination can drive an increase in frequencies of antibiotic resistance among nonvaccine serotypes of Streptococcus pneumoniae. Proc Natl Acad Sci USA 115(12):3102–3107. CrossRefPubMedGoogle Scholar
  16. 16.
    Commichau FM, Heidemann JL, Ficner R, Stulke J (2018) Making and breaking of an essential poison: the cyclases and phosphodiesterases that produce and degrade the essential second messenger cyclic di-AMP in bacteria. J Bacteriol.
  17. 17.
    Corrigan RM, Grundling A (2013) Cyclic di-AMP: another second messenger enters the fray. Nat Rev Microbiol 11(8):513–524. CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Bai Y, Yang J, Eisele LE, Underwood AJ, Koestler BJ, Waters CM, Metzger DW, Bai G (2013) Two DHH subfamily 1 proteins in Streptococcus pneumoniae possess cyclic di-AMP phosphodiesterase activity and affect bacterial growth and virulence. J Bacteriol 195(22):5123–5132. CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Zarrella TM, Metzger DW, Bai G (2018) Stress suppressor screening leads to detection of regulation of cyclic di-AMP homeostasis by a Trk family effector protein in Streptococcus pneumoniae. J Bacteriol 200:e00045-18. CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Pham TH, Liang ZX, Marcellin E, Turner MS (2016) Replenishing the cyclic-di-AMP pool: regulation of diadenylate cyclase activity in bacteria. Curr Genet 62(4):731–738. CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Gundlach J, Herzberg C, Kaever V, Gunka K, Hoffmann T, Weiss M, Gibhardt J, Thurmer A, Hertel D, Daniel R, Bremer E, Commichau FM, Stulke J (2017) Control of potassium homeostasis is an essential function of the second messenger cyclic di-AMP in Bacillus subtilis. Sci Signal 10(475):eaal3011. CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Mehne FM, Gunka K, Eilers H, Herzberg C, Kaever V, Stulke J (2013) Cyclic di-AMP homeostasis in Bacillus subtilis: both lack and high level accumulation of the nucleotide are detrimental for cell growth. J Biol Chem 288(3):2004–2017. CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Rismondo J, Gibhardt J, Rosenberg J, Kaever V, Halbedel S, Commichau FM (2015) Phenotypes associated with the essential diadenylate cyclase CdaA and its potential regulator CdaR in the human pathogen Listeria monocytogenes. J Bacteriol 198(3):416–426. CrossRefPubMedGoogle Scholar
  24. 24.
    Zhu Y, Pham TH, Nhiep TH, Vu NM, Marcellin E, Chakrabortti A, Wang Y, Waanders J, Lo R, Huston WM, Bansal N, Nielsen LK, Liang ZX, Turner MS (2016) Cyclic-di-AMP synthesis by the diadenylate cyclase CdaA is modulated by the peptidoglycan biosynthesis enzyme GlmM in Lactococcus lactis. Mol Microbiol 99(6):1015–1027. CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Bowman L, Zeden MS, Schuster CF, Kaever V, Grundling A (2016) New insights into the cyclic di-adenosine monophosphate (c-di-AMP) degradation pathway and the requirement of the cyclic dinucleotide for acid stress resistance in Staphylococcus aureus. J Biol Chem 291(53):26970–26986. CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Mengin-Lecreulx D, van Heijenoort J (1996) Characterization of the essential gene glmM encoding phosphoglucosamine mutase in Escherichia coli. J Biol Chem 271(1):32–39CrossRefGoogle Scholar
  27. 27.
    Shimazu K, Takahashi Y, Uchikawa Y, Shimazu Y, Yajima A, Takashima E, Aoba T, Konishi K (2008) Identification of the Streptococcus gordonii glmM gene encoding phosphoglucosamine mutase and its role in bacterial cell morphology, biofilm formation, and sensitivity to antibiotics. FEMS Immunol Med Microbiol 53(2):166–177. CrossRefPubMedGoogle Scholar
  28. 28.
    Gundlach J, Mehne FM, Herzberg C, Kampf J, Valerius O, Kaever V, Stulke J (2015) An essential poison: synthesis and degradation of cyclic di-AMP in Bacillus subtilis. J Bacteriol 197(20):3265–3274. CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Molzen TE, Burghout P, Bootsma HJ, Brandt CT, van der Gaast-de Jongh CE, Eleveld MJ, Verbeek MM, Frimodt-Moller N, Ostergaard C, Hermans PW (2011) Genome-wide identification of Streptococcus pneumoniae genes essential for bacterial replication during experimental meningitis. Infect Immun 79(1):288–297. CrossRefPubMedGoogle Scholar
  30. 30.
    Cron LE, Stol K, Burghout P, van Selm S, Simonetti ER, Bootsma HJ, Hermans PW (2011) Two DHH subfamily 1 proteins contribute to pneumococcal virulence and confer protection against pneumococcal disease. Infect Immun 79(9):3697–3710. CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Woodward JJ, Iavarone AT, Portnoy DA (2010) c-di-AMP secreted by intracellular Listeria monocytogenes activates a host type I interferon response. Science 328(5986):1703–1705. CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Kaplan Zeevi M, Shafir NS, Shaham S, Friedman S, Sigal N, Nir Paz R, Boneca IG, Herskovits AA (2013) Listeria monocytogenes multidrug resistance transporters and cyclic di-AMP, which contribute to type I interferon induction, play a role in cell wall stress. J Bacteriol 195(23):5250–5261. CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    Barker JR, Koestler BJ, Carpenter VK, Burdette DL, Waters CM, Vance RE, Valdivia RH (2013) STING-dependent recognition of cyclic di-AMP mediates type I interferon responses during Chlamydia trachomatis infection. MBio 4(3):e00018–e00013. CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Sauer JD, Sotelo-Troha K, von Moltke J, Monroe KM, Rae CS, Brubaker SW, Hyodo M, Hayakawa Y, Woodward JJ, Portnoy DA, Vance RE (2011) The N-ethyl-N-nitrosourea-induced Goldenticket mouse mutant reveals an essential function of Sting in the in vivo interferon response to Listeria monocytogenes and cyclic dinucleotides. Infect Immun 79(2):688–694. CrossRefPubMedGoogle Scholar
  35. 35.
    Parvatiyar K, Zhang Z, Teles RM, Ouyang S, Jiang Y, Iyer SS, Zaver SA, Schenk M, Zeng S, Zhong W, Liu ZJ, Modlin RL, Liu YJ, Cheng G (2012) The helicase DDX41 recognizes the bacterial secondary messengers cyclic di-GMP and cyclic di-AMP to activate a type I interferon immune response. Nat Immunol 13(12):1155–1161. CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Bowie AG (2012) Innate sensing of bacterial cyclic dinucleotides: more than just STING. Nat Immunol 13(12):1137–1139. CrossRefPubMedGoogle Scholar
  37. 37.
    Yamamoto T, Hara H, Tsuchiya K, Sakai S, Fang R, Matsuura M, Nomura T, Sato F, Mitsuyama M, Kawamura I (2012) Listeria monocytogenes strain-specific impairment of the TetR regulator underlies the drastic increase in cyclic di-AMP secretion and beta interferon-inducing ability. Infect Immun 80(7):2323–2332. CrossRefPubMedPubMedCentralGoogle Scholar
  38. 38.
    Corrigan RM, Abbott JC, Burhenne H, Kaever V, Grundling A (2011) c-di-AMP is a new second messenger in Staphylococcus aureus with a role in controlling cell size and envelope stress. PLoS Pathog 7(9):e1002217. CrossRefPubMedPubMedCentralGoogle Scholar
  39. 39.
    Luo Y, Helmann JD (2012) Analysis of the role of Bacillus subtilis sigma(M) in beta-lactam resistance reveals an essential role for c-di-AMP in peptidoglycan homeostasis. Mol Microbiol 83(3):623–639. CrossRefPubMedPubMedCentralGoogle Scholar
  40. 40.
    Oppenheimer-Shaanan Y, Wexselblatt E, Katzhendler J, Yavin E, Ben-Yehuda S (2011) c-di-AMP reports DNA integrity during sporulation in Bacillus subtilis. EMBO Rep 12(6):594–601. CrossRefPubMedPubMedCentralGoogle Scholar
  41. 41.
    Malli R, Epstein W (1998) Expression of the Kdp ATPase is consistent with regulation by turgor pressure. J Bacteriol 180(19):5102–5108CrossRefGoogle Scholar
  42. 42.
    Meury J, Robin A, Monnier-Champeix P (1985) Turgor-controlled K+ fluxes and their pathways in Escherichia coli. Eur J Biochem 151(3):613–619CrossRefGoogle Scholar
  43. 43.
    Epstein W (1986) Osmoregulation by potassium transport in Escherichia coli. FEMS Microbiol Lett 39:73–78CrossRefGoogle Scholar
  44. 44.
    Dinnbier U, Limpinsel E, Schmid R, Bakker EP (1988) Transient accumulation of potassium glutamate and its replacement by trehalose during adaptation of growing cells of Escherichia coli K-12 to elevated sodium chloride concentrations. Arch Microbiol 150(4):348–357CrossRefGoogle Scholar
  45. 45.
    Holtmann G, Bakker EP, Uozumi N, Bremer E (2003) KtrAB and KtrCD: two K+ uptake systems in Bacillus subtilis and their role in adaptation to hypertonicity. J Bacteriol 185(4):1289–1298CrossRefGoogle Scholar
  46. 46.
    Gries CM, Bose JL, Nuxoll AS, Fey PD, Bayles KW (2013) The Ktr potassium transport system in Staphylococcus aureus and its role in cell physiology, antimicrobial resistance and pathogenesis. Mol Microbiol 89(4):760–773. CrossRefPubMedPubMedCentralGoogle Scholar
  47. 47.
    Csonka LN (1989) Physiological and genetic responses of bacteria to osmotic stress. Microbiol Rev 53(1):121–147CrossRefGoogle Scholar
  48. 48.
    Ferguson GP, McLaggan D, Booth IR (1995) Potassium channel activation by glutathione-S-conjugates in Escherichia coli: protection against methylglyoxal is mediated by cytoplasmic acidification. Mol Microbiol 17(6):1025–1033CrossRefGoogle Scholar
  49. 49.
    Castaneda-Garcia A, Do TT, Blazquez J (2011) The K+ uptake regulator TrkA controls membrane potential, pH homeostasis and multidrug susceptibility in Mycobacterium smegmatis. J Antimicrob Chemother 66(7):1489–1498. CrossRefPubMedGoogle Scholar
  50. 50.
    Kashket ER, Barker SL (1977) Effects of potassium ions on the electrical and pH gradients across the membrane of Streptococcus lactis cells. J Bacteriol 130(3):1017–1023CrossRefGoogle Scholar
  51. 51.
    Hughes FM Jr, Cidlowski JA (1999) Potassium is a critical regulator of apoptotic enzymes in vitro and in vivo. Adv Enzym Regul 39:157–171CrossRefGoogle Scholar
  52. 52.
    Vasak M, Schnabl J (2016) Sodium and potassium ions in proteins and enzyme catalysis. Met Ions Life Sci 16:259–290. CrossRefPubMedGoogle Scholar
  53. 53.
    Tholema N, Bakker EP, Suzuki A, Nakamura T (1999) Change to alanine of one out of four selectivity filter glycines in KtrB causes a two orders of magnitude decrease in the affinities for both K+ and Na+ of the Na+ dependent K+ uptake system KtrAB from Vibrio alginolyticus. FEBS Lett 450(3):217–220CrossRefGoogle Scholar
  54. 54.
    Hanelt I, Tholema N, Kroning N, Vor der Bruggen M, Wunnicke D, Bakker EP (2011) KtrB, a member of the superfamily of K+ transporters. Eur J Cell Biol 90(9):696–704. CrossRefPubMedGoogle Scholar
  55. 55.
    Diskowski M, Mikusevic V, Stock C, Hanelt I (2015) Functional diversity of the superfamily of K(+) transporters to meet various requirements. Biol Chem 396(9–10):1003–1014. CrossRefPubMedGoogle Scholar
  56. 56.
    Altendorf K, Epstein W (1993) Kdp-ATPase of Escherichia coli. Cell Physiol Biochem 4:160–168CrossRefGoogle Scholar
  57. 57.
    Altendorf K, Voelkner P, Puppe W (1994) The sensor kinase KdpD and the response regulator KdpE control expression of the kdpFABC operon in Escherichia coli. Res Microbiol 145(5–6):374–381CrossRefGoogle Scholar
  58. 58.
    Alahari A, Ballal A, Apte SK (2001) Regulation of potassium-dependent Kdp-ATPase expression in the nitrogen-fixing cyanobacterium Anabaena torulosa. J Bacteriol 183(19):5778–5781. CrossRefPubMedPubMedCentralGoogle Scholar
  59. 59.
    Bai Y, Yang J, Zarrella TM, Zhang Y, Metzger DW, Bai G (2014) Cyclic di-AMP impairs potassium uptake mediated by a cyclic di-AMP binding protein in Streptococcus pneumoniae. J Bacteriol 196(3):614–623. CrossRefPubMedPubMedCentralGoogle Scholar
  60. 60.
    Corrigan RM, Campeotto I, Jeganathan T, Roelofs KG, Lee VT, Grundling A (2013) Systematic identification of conserved bacterial c-di-AMP receptor proteins. Proc Natl Acad Sci USA 110(22):9084–9089. CrossRefPubMedPubMedCentralGoogle Scholar
  61. 61.
    Kim H, Youn SJ, Kim SO, Ko J, Lee JO, Choi BS (2015) Structural studies of potassium transport protein KtrA regulator of conductance of K+ (RCK) C domain in complex with cyclic diadenosine monophosphate (c-di-AMP). J Biol Chem 290:16393–16402. CrossRefPubMedPubMedCentralGoogle Scholar
  62. 62.
    Moscoso JA, Schramke H, Zhang Y, Tosi T, Dehbi A, Jung K, Grundling A (2016) Binding of cyclic di-AMP to the Staphylococcus aureus sensor kinase KdpD occurs via the universal stress protein domain and downregulates the expression of the Kdp potassium transporter. J Bacteriol 198(1):98–110. CrossRefGoogle Scholar
  63. 63.
    Albright RA, Ibar JL, Kim CU, Gruner SM, Morais-Cabral JH (2006) The RCK domain of the KtrAB K+ transporter: multiple conformations of an octameric ring. Cell 126(6):1147–1159. CrossRefPubMedGoogle Scholar
  64. 64.
    Underwood AJ, Zhang Y, Metzger DW, Bai G (2014) Detection of cyclic di-AMP using a competitive ELISA with a unique pneumococcal cyclic di-AMP binding protein. J Microbiol Methods 107:58–62. CrossRefPubMedPubMedCentralGoogle Scholar
  65. 65.
    Epstein W (2003) The roles and regulation of potassium in bacteria. Prog Nucleic Acid Res Mol Biol 75:293–320CrossRefGoogle Scholar
  66. 66.
    Commichau FM, Stulke J (2018) Coping with an essential poison: a genetic suppressor analysis corroborates a key function of c-di-AMP in controlling potassium ion homeostasis in Gram-positive bacteria. J Bacteriol.
  67. 67.
    Smith WM, Pham TH, Lei L, Dou J, Soomro AH, Beatson SA, Dykes GA, Turner MS (2012) Heat resistance and salt hypersensitivity in Lactococcus lactis due to spontaneous mutation of llmg_1816 (gdpP) induced by high-temperature growth. Appl Environ Microbiol 78(21):7753–7759. CrossRefPubMedPubMedCentralGoogle Scholar
  68. 68.
    Whiteley AT, Pollock AJ, Portnoy DA (2015) The PAMP c-di-AMP is essential for Listeria monocytogenes growth in rich but not minimal media due to a toxic increase in (p)ppGpp. Cell Host Microbe 17(6):788–798. CrossRefPubMedPubMedCentralGoogle Scholar
  69. 69.
    Corrigan RM, Bowman L, Willis AR, Kaever V, Grundling A (2015) Cross-talk between two nucleotide-signaling pathways in Staphylococcus aureus. J Biol Chem 290(9):5826–5839. CrossRefPubMedPubMedCentralGoogle Scholar
  70. 70.
    Rao F, See RY, Zhang D, Toh DC, Ji Q, Liang ZX (2010) YybT is a signaling protein that contains a cyclic dinucleotide phosphodiesterase domain and a GGDEF domain with ATPase activity. J Biol Chem 285(1):473–482. CrossRefPubMedPubMedCentralGoogle Scholar
  71. 71.
    Tadmor K, Pozniak Y, Burg Golani T, Lobel L, Brenner M, Sigal N, Herskovits AA (2014) Listeria monocytogenes MDR transporters are involved in LTA synthesis and triggering of innate immunity during infection. Front Cell Infect Microbiol 4:16. CrossRefPubMedPubMedCentralGoogle Scholar
  72. 72.
    Schwartz KT, Carleton JD, Quillin SJ, Rollins SD, Portnoy DA, Leber JH (2012) Hyperinduction of host beta interferon by a Listeria monocytogenes strain naturally overexpressing the multidrug efflux pump MdrT. Infect Immun 80(4):1537–1545. CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Nature Switzerland AG 2020

Authors and Affiliations

  1. 1.Laboratory of Molecular Biology, Center for Cancer ResearchNational Cancer InstituteBethesdaUSA
  2. 2.Department of Immunology and Microbial DiseaseAlbany Medical CollegeAlbanyUSA

Personalised recommendations