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Mycopathologia

, Volume 178, Issue 5–6, pp 341–348 | Cite as

Calcineurin-Mediated Regulation of Hyphal Growth, Septation, and Virulence in Aspergillus fumigatus

  • Praveen R. JuvvadiEmail author
  • Frédéric Lamoth
  • William J. SteinbachEmail author
Article

Abstract

Calcineurin is a heterodimeric protein phosphatase complex composed of catalytic (CnaA) and regulatory (CnaB) subunits and plays diverse roles in regulating fungal stress responses, morphogenesis, and pathogenesis. Fungal pathogens utilize the calcineurin pathway to survive in the host environment and cause life-threatening infections. The immunosuppressive calcineurin inhibitors (FK506 and cyclosporine A) are active against fungi, making calcineurin a promising antifungal drug target. Here, we review novel findings on calcineurin localization and functions in Aspergillus fumigatus hyphal growth and septum formation through regulation of proteins involved in cell wall biosynthesis. Extensive mutational analysis in the functional domains of A. fumigatus CnaA has led to an understanding of the relevance of these domains for the localization and function of CnaA at the hyphal septum. An evolutionarily conserved novel mode of calcineurin regulation by phosphorylation in filamentous fungi was found to be responsible for virulence in A. fumigatus. This finding of a filamentous fungal-specific mechanism controlling hyphal growth and virulence represents a potential target for antifungal therapy.

Keywords

Calcineurin Aspergillus fumigatus Hyphal growth Septum Virulence 

Notes

Acknowledgments

These studies were funded by NIH/NIAID 1 R56 AI077648-01A2 and 1 R21 AI097541-01A1 Grants to WJS. FL is supported by the Swiss Foundation for Medical-Biological Grants and the Swiss National Science Foundation (P3SMP3-151742).

References

  1. 1.
    Kontoyiannis DP, Marr KA, Park BJ, Alexander BD, Anaissie EJ, Walsh TJ, et al. Prospective surveillance for invasive fungal infections in hematopoietic stem cell transplant recipients, 2001–2006: overview of the transplant-associated infection surveillance network (TRANSNET) database. Clin Infect Dis. 2010;50(8):1091–100.PubMedCrossRefGoogle Scholar
  2. 2.
    Steinbach WJ, Marr KA, Anaissie EJ, Azie N, Quan S-P, Meier-Kriesche H-U, et al. Clinical epidemiology of 960 patients with invasive aspergillosis from the PATH Alliance registry. J Infect. 2012;65(5):453–64.PubMedCrossRefGoogle Scholar
  3. 3.
    Steinbach WJ, Reedy JL, Cramer RA, Perfect JR, Heitman J. Harnessing calcineurin as a novel anti-infective agent against invasive fungal infections. Nat Rev Microbiol. 2007;5(6):418–30.PubMedCrossRefGoogle Scholar
  4. 4.
    Ho S, Clipstone N, Timmermann L, Northrop J, Graef I, Fiorentino D, et al. The Mechanism of Action of Cyclosporin A and FK506. Clin Immunol Immunopathol. 1996;80(3):S40–5.PubMedCrossRefGoogle Scholar
  5. 5.
    Stewart AA, Ingebritsen TS, Manalan A, Klee CB, Cohen P. Discovery of A Ca2+-and calmodulin-dependent protein phosphatase. FEBS Lett. 1982;137(1):80–4.PubMedCrossRefGoogle Scholar
  6. 6.
    Cyert MS. Genetic analysis of calmodulin and its targets in Saccharomyces cerevisiae. Annu Rev Genet. 2001;35(1):647–72.PubMedCrossRefGoogle Scholar
  7. 7.
    Sugiura R, Sio SO, Shuntoh H, Kuno T. Calcineurin phosphatase in signal transduction: lessons from fission yeast. Genes Cells. 2002;7(7):619–27.PubMedCrossRefGoogle Scholar
  8. 8.
    Fox DS, Heitman J. Good fungi gone bad: the corruption of calcineurin. BioEssays. 2002;24(10):894–903.PubMedCrossRefGoogle Scholar
  9. 9.
    Hemenway C, Heitman J. Calcineurin. Cell Biochem Biophys. 1999;30(1):115–51.PubMedCrossRefGoogle Scholar
  10. 10.
    Aramburu J, Rao A, Klee CB. Calcineurin: from structure to function. Curr Top Cell Regul. 2000;36:237–95.PubMedCrossRefGoogle Scholar
  11. 11.
    Rusnak F, Mertz P. Calcineurin: form and Function. Physiol Rev. 2000;80(4):1483–521.PubMedGoogle Scholar
  12. 12.
    Cyert MS, Kunisawa R, Kaim D, Thorner J. Yeast has homologs (CNA1 and CNA2 gene products) of mammalian calcineurin, a calmodulin-regulated phosphoprotein phosphatase. Proc Natl Acad Sci USA. 1991;88(16):7376–80.PubMedCentralPubMedCrossRefGoogle Scholar
  13. 13.
    Yoshida T, Toda T, Yanagida M. A calcineurin-like gene ppb1 + in fission yeast: mutant defects in cytokinesis, cell polarity, mating and spindle pole body positioning. J Cell Sci. 1994;107(7):1725–35.PubMedGoogle Scholar
  14. 14.
    Fox DS, Cruz MC, Sia RAL, Ke H, Cox GM, Cardenas ME, et al. Calcineurin regulatory subunit is essential for virulence and mediates interactions with FKBP12–FK506 in Cryptococcus neoformans. Mol Microbiol. 2001;39(4):835–49.PubMedCrossRefGoogle Scholar
  15. 15.
    Kraus PR, Heitman J. Coping with stress: calmodulin and calcineurin in model and pathogenic fungi. Biochem Biophys Res Commun. 2003;311(4):1151–7.PubMedCrossRefGoogle Scholar
  16. 16.
    Cruz MC, Goldstein AL, Blankenship JR, Del Poeta M, Davis D, Cardenas ME, et al. Calcineurin is essential for survival during membrane stress in Candida albicans. EMBO J. 2002;21(4):546–59.PubMedCentralPubMedCrossRefGoogle Scholar
  17. 17.
    Bader T, Bodendorfer B, Schroppel K, Morschhauser J. Calcineurin Is Essential for Virulence in Candida albicans. Infect Immun. 2003;71(9):5344–54.PubMedCentralPubMedCrossRefGoogle Scholar
  18. 18.
    Reedy JL, Filler SG, Heitman J. Elucidating the Candida albicans calcineurin signaling cascade controlling stress response and virulence. Fungal Genet Biol. 2010;47(2):107–16.PubMedCentralPubMedCrossRefGoogle Scholar
  19. 19.
    Chen YL, Kozubowski L, Cardenas ME, Heitman J. On the roles of calcineurin in fungal growth and pathogenesis. Curr Fungal Infect Rep. 2010;4:244–55.CrossRefGoogle Scholar
  20. 20.
    Odom A, Muir S, Lim E, Toffaletti DL, Perfect J, Heitman J. Calcineurin is required for virulence of Cryptococcus neoformans. EMBO J. 1997;16(10):2576–89.PubMedCentralPubMedCrossRefGoogle Scholar
  21. 21.
    Zhang Y, Sugiura R, Lu Y, Asami M, Maeda T, Itoh T, et al. Phosphatidylinositol 4-phosphate 5-kinase Its3 and calcineurin Ppb1 coordinately regulate cytokinesis in fission yeast. J Biol Chem. 2000;275(45):35600–6.PubMedCrossRefGoogle Scholar
  22. 22.
    Kita A, Sugiura R, Shoji H, He Y, Deng L, Lu Y, et al. Loss of Apm1, the μ1 subunit of the clathrin-associated adaptor-protein-1 complex, causes distinct phenotypes and synthetic lethality with calcineurin deletion in fission yeast. Mol Biol Cell. 2004;15(6):2920–31.PubMedCentralPubMedCrossRefGoogle Scholar
  23. 23.
    Rasmussen C, Garen C, Brining S, Kincaid RL, Means RL, Means AR. The calmodulin-dependent protein phosphatase catalytic subunit (calcineurin A) is an essential gene in Aspergillus nidulans. EMBO J. 1994;13(16):3917–24.PubMedCentralPubMedGoogle Scholar
  24. 24.
    Prokisch H, Yarden O, Dieminger M, Tropschug M, Barthelmess IB. Impairment of calcineurin function in Neurospora crassa reveals its essential role in hyphal growth, morphology and maintenance of the apical Ca2+ gradient. Mol Gen Genet. 1997;256:104–14.PubMedCrossRefGoogle Scholar
  25. 25.
    Juvvadi PR, Kuroki Y, Arioka M, Nakajima H, Kitamoto K. Functional analysis of the calcineurin-encoding gene cnaA from Aspergillus oryzae: evidence for its putative role in stress adaptation. Arch Microbiol. 2003;179(6):416–22.PubMedGoogle Scholar
  26. 26.
    Harel A, Bercovich S, Yarden O. Calcineurin Is Required for Sclerotial Development and Pathogenicity of Sclerotinia sclerotiorum in an Oxalic Acid-Independent Manner. Mol Plant Microbe Interact. 2006;19(6):682–93. doi: 10.1094/mpmi-19-0682.PubMedCrossRefGoogle Scholar
  27. 27.
    Choi J, Kim Y, Kim S, Park J, Lee Y-H. MoCRZ1, a gene encoding a calcineurin-responsive transcription factor, regulates fungal growth and pathogenicity of Magnaporthe oryzae. Fungal Genet Biol. 2009;46(3):243–54.PubMedCrossRefGoogle Scholar
  28. 28.
    Liu J, Farmer JD Jr, Lane WS, Friedman J, Weissman I, Schreiber SL. Calcineurin is a common target of cyclophilin-cyclosporin A and FKBP-FK506 complexes. Cell. 1991;66(4):807–15.PubMedCrossRefGoogle Scholar
  29. 29.
    Blankenship JR, Wormley FL, Boyce MK, Schell WA, Filler SG, Perfect JR, et al. Calcineurin Is Essential for Candida albicans Survival in Serum and Virulence. Eukaryot Cell. 2003;2(3):422–30. doi: 10.1128/ec.2.3.422-430.2003.PubMedCentralPubMedCrossRefGoogle Scholar
  30. 30.
    Bader T, Schröppel K, Bentink S, Agabian N, Köhler G, Morschhäuser J. Role of calcineurin in stress resistance, morphogenesis, and virulence of a candida albicans wild-type strain. Infect Immun. 2006;74(7):4366–9.PubMedCentralPubMedCrossRefGoogle Scholar
  31. 31.
    Steinbach WJ, Cramer RA Jr, Perfect BZ, Asfaw YG, Sauer TC, Najvar LK, et al. Calcineurin controls growth, morphology, and pathogenicity in Aspergillus fumigatus. Eukaryot Cell. 2006;5(7):1091–103.PubMedCentralPubMedCrossRefGoogle Scholar
  32. 32.
    Juvvadi PR, Fortwendel JR, Rogg LE, Burns KA, Randell SH, Steinbach WJ. Localization and activity of the calcineurin catalytic and regulatory subunit complex at the septum is essential for hyphal elongation and proper septation in Aspergillus fumigatus. Mol Microbiol. 2011;82(5):1235–59.PubMedCentralPubMedCrossRefGoogle Scholar
  33. 33.
    Garrett-Engele P, Moilanen B, Cyert M. Calcineurin, the Ca2+/calmodulin-dependent protein phosphatase, is essential in yeast mutants with cell integrity defects and in mutants that lack a functional vacuolar H(+)-ATPase. Mol Cell Biol. 1995;15(8):4103–14.PubMedCentralPubMedGoogle Scholar
  34. 34.
    Delgado-Jarana J, Sousa S, Gonzalez F, Rey M, Llobell A. ThHog1 controls the hyperosmotic stress response in Trichoderma harzianum. Microbiology. 2006;152(6):1687–700.PubMedCrossRefGoogle Scholar
  35. 35.
    Fortwendel JR, Juvvadi PR, Pinchai N, Perfect BZ, Alspaugh JA, Perfect JR, et al. Differential effects of inhibiting chitin and 1,3-{beta}-D-glucan synthesis in ras and calcineurin mutants of Aspergillus fumigatus. Antimicrob Agents Chemother. 2009;53(2):476–82.PubMedCentralPubMedCrossRefGoogle Scholar
  36. 36.
    Cramer RA Jr, Perfect BZ, Pinchai N, Park S, Perlin DS, Asfaw YG, et al. Calcineurin target CrzA regulates conidial germination, hyphal growth, and pathogenesis of Aspergillus fumigatus. Eukaryot Cell. 2008;7(7):1085–97.PubMedCentralPubMedCrossRefGoogle Scholar
  37. 37.
    Juvvadi PR, Fortwendel JR, Pinchai N, Perfect BZ, Heitman J, Steinbach WJ. Calcineurin localizes to the hyphal septum in Aspergillus fumigatus: implications for septum formation and conidiophore development. Eukaryot Cell. 2008;7(9):1606–10.PubMedCentralPubMedCrossRefGoogle Scholar
  38. 38.
    Juvvadi PR, Gehrke C, Fortwendel JR, Lamoth F, Soderblom EJ, Cook EC, et al. Phosphorylation of calcineurin at a novel serine-proline rich region orchestrates hyphal growth and virulence in Aspergillus fumigatus. PLoS Pathog. 2013;9(8):e1003564.PubMedCentralPubMedCrossRefGoogle Scholar
  39. 39.
    Li H, Zhang L, Rao A, Harrison SC, Hogan PG. Structure of calcineurin in complex with pvivit peptide: portrait of a low-affinity signalling interaction. J Mol Biol. 2007;369(5):1296–306.PubMedCrossRefGoogle Scholar
  40. 40.
    Roy J, Li H, Hogan PG, Cyert MS. A conserved docking site modulates substrate affinity for calcineurin, signaling output, and in vivo function. Mol Cell. 2007;25(6):889–901.PubMedCentralPubMedCrossRefGoogle Scholar
  41. 41.
    Ye Q, Wang H, Zheng J, Wei Q, Jia Z. The complex structure of calmodulin bound to a calcineurin peptide. Proteins Struct Funct Bioinform. 2008;73(1):19–27.CrossRefGoogle Scholar
  42. 42.
    Chen S, Song Y, Cao J, Wang G, Wei H, Xu X, et al. Localization and function of calmodulin in live-cells of Aspergillus nidulans. Fungal Genet Biol. 2010;47(3):268–78.PubMedCrossRefGoogle Scholar
  43. 43.
    Jiang B, Cyert MS. Identification of a novel region critical for calcineurin function in vivo and in vitro. J Biol Chem. 1999;274(26):18543–51.PubMedCrossRefGoogle Scholar
  44. 44.
    Berzal S, Alique M, Ruiz-Ortega M, Egido J, Ortiz A, Ramos AM. GSK3, snail, and adhesion molecule regulation by cyclosporine a in renal tubular cells. Toxicol Sci. 2012;127(2):425–37.PubMedCrossRefGoogle Scholar
  45. 45.
    Singh TJ, Wang JH. Phosphorylation of calcineurin by glycogen synthase (casein) kinase-1. Biochem Cell Biol. 1987;65(10):917–21.PubMedCrossRefGoogle Scholar
  46. 46.
    Hashimoto Y, Soderling TR. Regulation of calcineurin by phosphorylation. Identification of the regulatory site phosphorylated by Ca2+/calmodulin-dependent protein kinase II and protein kinase C. J Biol Chem. 1989;264(28):16524–9.PubMedGoogle Scholar
  47. 47.
    Martensen TM, Martin BM, Kincaid RL. Identification of the site on calcineurin phosphorylated by calcium/CaM-dependent kinase II: modification of the CaM-binding domain. Biochemistry. 1989;28(24):9243–7.PubMedCrossRefGoogle Scholar
  48. 48.
    Hashimoto Y, King MM, Soderling TR. Regulatory interactions of calmodulin-binding proteins: phosphorylation of calcineurin by autophosphorylated Ca2+/calmodulin-dependent protein kinase II. Proc Natl Acad Sci. 1988;85(18):7001–5.PubMedCentralPubMedCrossRefGoogle Scholar
  49. 49.
    Kume K, Koyano T, Kanai M, Toda T, Hirata D. Calcineurin ensures a link between the DNA replication checkpoint and microtubule-dependent polarized growth. Nat Cell Biol. 2011;13(3):234–42.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  1. 1.Division of Pediatric Infectious Diseases, Department of PediatricsDuke University Medical CenterDurhamUSA
  2. 2.Infectious Diseases Service, Department of MedicineLausanne University HospitalLausanneSwitzerland
  3. 3.Institute of MicrobiologyLausanne University HospitalLausanneSwitzerland
  4. 4.Department of Molecular Genetics and MicrobiologyDuke University Medical CenterDurhamUSA

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