A Single Protein S-acyl Transferase Acts through Diverse Substrates to Determine Cryptococcal Morphology, Stress Tolerance, and Pathogenic Outcome

English version České info

Cryptococcus neoformans is a ubiquitous environmental yeast that kills over 625,000 people annually, mainly in developing countries. Healthy humans frequently inhale infectious particles without noticeable symptoms. However, in immunocompromised people, the initial lung infection can spread to other sites, particularly to the central nervous system where it causes lethal brain infection. The infected host responds by deploying immune cells to engulf and kill the yeast, but C. neoformans can survive this engulfment and even multiply within the host cells. To understand the interactions between the invading microbe and host cells we screened 1,201 fungal mutants to identify fungal factors that influence these processes. One mutant, lacking an enzyme that modifies proteins with the lipid palmitate, showed an increase in engulfment by the host along with dramatic defects in morphology, stress resistance, and virulence. We went on to identify the proteins this enzyme modifies and explain how its absence leads to altered cell wall synthesis, signal transduction, and membrane trafficking; these changes explain the behavior of the mutant. We also found that the mutant could not cause disease in an animal model. Our work shows that protein palmitoylation is critical for cryptococcal pathogenesis and presents a potential avenue for antifungal therapy.

Vyšlo v časopise: A Single Protein S-acyl Transferase Acts through Diverse Substrates to Determine Cryptococcal Morphology, Stress Tolerance, and Pathogenic Outcome. PLoS Pathog 11(5): e32767. doi:10.1371/journal.ppat.1004908
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1004908

Souhrn

Zdroje

1. Sabiiti W, May RC. Mechanisms of infection by the human fungal pathogen Cryptococcus neoformans. Future Microbiol. 2012;7(11):1297–313. Epub 2012/10/19. doi: 10.2217/fmb.12.102 23075448

2. Pyrgos V, Seitz AE, Steiner CA, Prevots DR, Williamson PR. Epidemiology of cryptococcal meningitis in the US: 1997–2009. PLoS One. 2013;8(2):e56269. Epub 2013/03/05. doi: 10.1371/journal.pone.0056269 23457543

3. Pappas PG. Cryptococcal infections in non-HIV-infected patients. Trans Am Clin Climatol Assoc. 2013;124:61–79. Epub 2013/07/23. PMCID: 3715903. 23874010

4. Bratton EW, El Husseini N, Chastain CA, Lee MS, Poole C, Sturmer T, et al. Comparison and temporal trends of three groups with cryptococcosis: HIV-infected, solid organ transplant, and HIV-negative/non-transplant. PLoS One. 2012;7(8):e43582. Epub 2012/09/01. doi: 10.1371/journal.pone.0043582 22937064

5. Mirza SA, Phelan M, Rimland D, Graviss E, Hamill R, Brandt ME, et al. The changing epidemiology of cryptococcosis: an update from population-based active surveillance in 2 large metropolitan areas, 1992–2000. Clin Infect Dis. 2003;36(6):789–94. Epub 2003/03/11. doi: 10.1086/368091 12627365

6. Wozniak KL, Hardison S, Olszewski M, Wormley FL Jr. Induction of protective immunity against cryptococcosis. Mycopathologia. 2012;173(5–6):387–94. Epub 2011/12/07. doi: 10.1007/s11046-011-9505-8 22354778

7. Alanio A, Desnos-Ollivier M, Dromer F. Dynamics of Cryptococcus neoformans-macrophage interactions reveal that fungal background influences outcome during cryptococcal meningoencephalitis in humans. MBio. 2011;2(4). Epub 2011/08/11. doi: 10.1128/mBio.00158-11

8. Sabiiti W, Robertson E, Beale MA, Johnston SA, Brouwer AE, Loyse A, et al. Efficient phagocytosis and laccase activity affect the outcome of HIV-associated cryptococcosis. J Clin Invest. 2014;124(5):2000–8. Epub 2014/04/20. doi: 10.1172/JCI72950 24743149

9. Srikanta D, Yang M, Williams M, Doering TL. A sensitive high-throughput assay for evaluating host-pathogen interactions in Cryptococcus neoformans infection. PLoS One. 2011;6(7):e22773. Epub 2011/08/11. doi: 10.1371/journal.pone.0022773 21829509

10. Liu OW, Chun CD, Chow ED, Chen C, Madhani HD, Noble SM. Systematic genetic analysis of virulence in the human fungal pathogen Cryptococcus neoformans. Cell. 2008;135(1):174–88. Epub 2008/10/16. doi: 10.1016/j.cell.2008.07.046 18854164

11. Linder ME, Jennings BC. Mechanism and function of DHHC S-acyltransferases. Biochem Soc Trans. 2013;41(1):29–34. Epub 2013/01/30. doi: 10.1042/BST20120328 23356254

12. Lobo S, Greentree WK, Linder ME, Deschenes RJ. Identification of a Ras palmitoyltransferase in Saccharomyces cerevisiae. J Biol Chem. 2002;277(43):41268–73. Epub 2002/08/24. doi: 10.1074/jbc.M206573200 12193598

13. Roth AF, Feng Y, Chen L, Davis NG. The yeast DHHC cysteine-rich domain protein Akr1p is a palmitoyl transferase. J Cell Biol. 2002;159(1):23–8. Epub 2002/10/09. doi: 10.1083/jcb.200206120 12370247

14. Veit M, Serebryakova MV, Kordyukova LV. Palmitoylation of influenza virus proteins. Biochem Soc Trans. 2013;41(1):50–5. Epub 2013/01/30. doi: 10.1042/BST20120210 23356257

15. Hicks SW, Charron G, Hang HC, Galan JE. Subcellular targeting of Salmonella virulence proteins by host-mediated S-palmitoylation. Cell Host Microbe. 2011;10(1):9–20. Epub 2011/07/20. doi: 10.1016/j.chom.2011.06.003 21767808

16. Beck JR, Fung C, Straub KW, Coppens I, Vashisht AA, Wohlschlegel JA, et al. A Toxoplasma palmitoyl acyl transferase and the palmitoylated armadillo repeat protein TgARO govern apical rhoptry tethering and reveal a critical role for the rhoptries in host cell invasion but not egress. PLoS Pathog. 2013;9(2):e1003162. Epub 2013/02/15. doi: 10.1371/journal.ppat.1003162 23408890

17. Frenal K, Tay CL, Mueller C, Bushell ES, Jia Y, Graindorge A, et al. Global analysis of apicomplexan protein S-acyl transferases reveals an enzyme essential for invasion. Traffic. 2013;14(8):895–911. Epub 2013/05/04. doi: 10.1111/tra.12081 23638681

18. Jones ML, Collins MO, Goulding D, Choudhary JS, Rayner JC. Analysis of protein palmitoylation reveals a pervasive role in Plasmodium development and pathogenesis. Cell Host Microbe. 2012;12(2):246–58. Epub 2012/08/21. doi: 10.1016/j.chom.2012.06.005 22901544

19. Nichols CB, Ferreyra J, Ballou ER, Alspaugh JA. Subcellular localization directs signaling specificity of the Cryptococcus neoformans Ras1 protein. Eukaryot Cell. 2009;8(2):181–9. Epub 2008/12/23. doi: 10.1128/EC.00351-08 19098128

20. Fortwendel JR, Juvvadi PR, Rogg LE, Asfaw YG, Burns KA, Randell SH, et al. Plasma membrane localization is required for RasA-mediated polarized morphogenesis and virulence of Aspergillus fumigatus. Eukaryot Cell. 2012;11(8):966–77. Epub 2012/05/09. doi: 10.1128/EC.00091-12 22562470

21. Piispanen AE, Bonnefoi O, Carden S, Deveau A, Bassilana M, Hogan DA. Roles of Ras1 membrane localization during Candida albicans hyphal growth and farnesol response. Eukaryot Cell. 2011;10(11):1473–84. Epub 2011/09/13. doi: 10.1128/EC.05153-11 21908593

22. McQuiston TJ, Williamson PR. Paradoxical roles of alveolar macrophages in the host response to Cryptococcus neoformans. J Infect Chemother. 2012;18(1):1–9. Epub 2011/11/03. doi: 10.1007/s10156-011-0306-2 22045161

23. Coelho C, Bocca AL, Casadevall A. The intracellular life of Cryptococcus neoformans. Annu Rev Pathol. 2014;9:219–38. Epub 2013/09/21. doi: 10.1146/annurev-pathol-012513-104653 24050625

24. O'Meara TR, Holmer SM, Selvig K, Dietrich F, Alspaugh JA. Cryptococcus neoformans Rim101 is associated with cell wall remodeling and evasion of the host immune responses. MBio. 2013;4(1). Epub 2013/01/17. doi: 10.1128/mBio.00522-12

25. Choi J, Vogl AW, Kronstad JW. Regulated expression of cyclic AMP-dependent protein kinase A reveals an influence on cell size and the secretion of virulence factors in Cryptococcus neoformans. Mol Microbiol. 2012;85(4):700–15. Epub 2012/06/22. doi: 10.1111/j.1365-2958.2012.08134.x 22717009

26. Kumar P, Heiss C, Santiago-Tirado FH, Black I, Azadi P, Doering TL. Pbx proteins in Cryptococcus neoformans cell wall remodeling and capsule assembly. Eukaryot Cell. 2014;13(5):560–71. Epub 2014/03/04. doi: 10.1128/EC.00290-13 24585882

27. Inglis DO, Skrzypek MS, Liaw E, Moktali V, Sherlock G, Stajich JE. Literature-based gene curation and proposed genetic nomenclature for Cryptococcus. Eukaryot Cell. 2014;13(7):878–83. Epub 2014/05/13. doi: 10.1128/EC.00083-14 24813190

28. Roth AF, Wan J, Bailey AO, Sun B, Kuchar JA, Green WN, et al. Global analysis of protein palmitoylation in yeast. Cell. 2006;125(5):1003–13. Epub 2006/06/06. doi: 10.1016/j.cell.2006.03.042 16751107

29. Nielsen K, Cox GM, Wang P, Toffaletti DL, Perfect JR, Heitman J. Sexual cycle of Cryptococcus neoformans var. grubii and virulence of congenic a and alpha isolates. Infect Immun. 2003;71(9):4831–41. Epub 2003/08/23. PMCID: 187335. 12933823

30. Wozniak KL, Levitz SM. Cryptococcus neoformans enters the endolysosomal pathway of dendritic cells and is killed by lysosomal components. Infect Immun. 2008;76(10):4764–71. Epub 2008/08/06. doi: 10.1128/IAI.00660-08 18678670

31. Smith LM, Dixon EF, May RC. The fungal pathogen Cryptococcus neoformans manipulates macrophage phagosome maturation. Cell Microbiol. 2014. Epub 2014/11/15. doi: 10.1111/cmi.12394

32. Banks IR, Specht CA, Donlin MJ, Gerik KJ, Levitz SM, Lodge JK. A chitin synthase and its regulator protein are critical for chitosan production and growth of the fungal pathogen Cryptococcus neoformans. Eukaryot Cell. 2005;4(11):1902–12. Epub 2005/11/10. doi: 10.1128/EC.4.11.1902-1912.2005 16278457

33. Reese AJ, Yoneda A, Breger JA, Beauvais A, Liu H, Griffith CL, et al. Loss of cell wall alpha(1–3) glucan affects Cryptococcus neoformans from ultrastructure to virulence. Mol Microbiol. 2007;63(5):1385–98. Epub 2007/01/25. doi: 10.1111/j.1365-2958.2006.05551.x 17244196

34. Kuranda K, Leberre V, Sokol S, Palamarczyk G, Francois J. Investigating the caffeine effects in the yeast Saccharomyces cerevisiae brings new insights into the connection between TOR, PKC and Ras/cAMP signalling pathways. Mol Microbiol. 2006;61(5):1147–66. Epub 2006/08/24. doi: 10.1111/j.1365-2958.2006.05300.x 16925551

35. Reese AJ, Doering TL. Cell wall alpha-1,3-glucan is required to anchor the Cryptococcus neoformans capsule. Mol Microbiol. 2003;50(4):1401–9. Epub 2003/11/19. 14622425

36. Johnston SA, May RC. Cryptococcus interactions with macrophages: evasion and manipulation of the phagosome by a fungal pathogen. Cell Microbiol. 2013;15(3):403–11. Epub 2012/11/07. doi: 10.1111/cmi.12067 23127124

37. Levitz SM, Nong SH, Seetoo KF, Harrison TS, Speizer RA, Simons ER. Cryptococcus neoformans resides in an acidic phagolysosome of human macrophages. Infect Immun. 1999;67(2):885–90. Epub 1999/01/23. PMCID: 96400. 9916104

38. Charron G, Zhang MM, Yount JS, Wilson J, Raghavan AS, Shamir E, et al. Robust fluorescent detection of protein fatty-acylation with chemical reporters. J Am Chem Soc. 2009;131(13):4967–75. Epub 2009/03/14. doi: 10.1021/ja810122f 19281244

39. Yount JS, Moltedo B, Yang YY, Charron G, Moran TM, Lopez CB, et al. Palmitoylome profiling reveals S-palmitoylation-dependent antiviral activity of IFITM3. Nat Chem Biol. 2010;6(8):610–4. Epub 2010/07/06. doi: 10.1038/nchembio.405 20601941

40. Yount JS, Zhang MM, Hang HC. Visualization and Identification of Fatty Acylated Proteins Using Chemical Reporters. Curr Protoc Chem Biol. 2011;3(2):65–79. Epub 2011/05/01. doi: 10.1002/9780470559277.ch100225 23061028

41. Zhang MM, Wu PY, Kelly FD, Nurse P, Hang HC. Quantitative control of protein S-palmitoylation regulates meiotic entry in fission yeast. PLoS Biol. 2013;11(7):e1001597. Epub 2013/07/12. doi: 10.1371/journal.pbio.1001597 23843742

42. Lam KK, Davey M, Sun B, Roth AF, Davis NG, Conibear E. Palmitoylation by the DHHC protein Pfa4 regulates the ER exit of Chs3. J Cell Biol. 2006;174(1):19–25. Epub 2006/07/05. doi: 10.1083/jcb.200602049 16818716

43. Gonzalez Montoro A, Chumpen Ramirez S, Quiroga R, Valdez Taubas J. Specificity of transmembrane protein palmitoylation in yeast. PLoS One. 2011;6(2):e16969. Epub 2011/03/09. doi: 10.1371/journal.pone.0016969 21383992

44. Sanchez-Mir L, Franco A, Martin-Garcia R, Madrid M, Vicente-Soler J, Soto T, et al. Rho2 palmitoylation is required for plasma membrane localization and proper signaling to the fission yeast cell integrity mitogen- activated protein kinase pathway. Mol Cell Biol. 2014;34(14):2745–59. Epub 2014/05/14. doi: 10.1128/MCB.01515-13 24820419

45. Young E, Zheng ZY, Wilkins AD, Jeong HT, Li M, Lichtarge O, et al. Regulation of Ras localization and cell transformation by evolutionarily conserved palmitoyltransferases. Mol Cell Biol. 2014;34(3):374–85. Epub 2013/11/20. doi: 10.1128/MCB.01248-13 24248599

46. Baker LG, Specht CA, Donlin MJ, Lodge JK. Chitosan, the deacetylated form of chitin, is necessary for cell wall integrity in Cryptococcus neoformans. Eukaryot Cell. 2007;6(5):855–67. Epub 2007/04/03. doi: 10.1128/EC.00399-06 17400891

47. Del Poeta M. Role of phagocytosis in the virulence of Cryptococcus neoformans. Eukaryot Cell. 2004;3(5):1067–75. Epub 2004/10/08. 10.1128/EC.3.5.1067-1075.2004. 15470235

48. Blanc M, Blaskovic S, van der Goot FG. Palmitoylation, pathogens and their host. Biochem Soc Trans. 2013;41(1):84–8. Epub 2013/01/30. doi: 10.1042/BST20120337 23356263

49. Goldston AM, Sharma AI, Paul KS, Engman DM. Acylation in trypanosomatids: an essential process and potential drug target. Trends Parasitol. 2014;30(7):350–60. Epub 2014/06/24. doi: 10.1016/j.pt.2014.05.003 24954795

50. Hole CR, Bui H, Wormley FL Jr., Wozniak KL. Mechanisms of dendritic cell lysosomal killing of Cryptococcus. Sci Rep. 2012;2:739. Epub 2012/10/18. doi: 10.1038/srep00739 23074646

51. Okagaki LH, Strain AK, Nielsen JN, Charlier C, Baltes NJ, Chretien F, et al. Cryptococcal cell morphology affects host cell interactions and pathogenicity. PLoS Pathog. 2010;6(6):e1000953. Epub 2010/06/30. doi: 10.1371/journal.ppat.1000953 20585559

52. Lam WC, Gerik KJ, Lodge JK. Role of Cryptococcus neoformans Rho1 GTPases in the PKC1 signaling pathway in response to thermal stress. Eukaryot Cell. 2013;12(1):118–31. Epub 2012/11/20. doi: 10.1128/EC.05305-11 23159519

53. Alspaugh JA, Perfect JR, Heitman J. Cryptococcus neoformans mating and virulence are regulated by the G-protein alpha subunit GPA1 and cAMP. Genes Dev. 1997;11(23):3206–17. Epub 1998/02/12. PMCID: 316752. 9389652

54. Levitz SM, Specht CA. The molecular basis for the immunogenicity of Cryptococcus neoformans mannoproteins. FEMS Yeast Res. 2006;6(4):513–24. Epub 2006/05/16. doi: 10.1111/j.1567-1364.2006.00071.x 16696647

55. Teixeira PA, Penha LL, Mendonca-Previato L, Previato JO. Mannoprotein MP84 mediates the adhesion of Cryptococcus neoformans to epithelial lung cells. Front Cell Infect Microbiol. 2014;4:106. Epub 2014/09/06. doi: 10.3389/fcimb.2014.00106 25191644

56. Bueter CL, Lee CK, Rathinam VA, Healy GJ, Taron CH, Specht CA, et al. Chitosan but not chitin activates the inflammasome by a mechanism dependent upon phagocytosis. J Biol Chem. 2011;286(41):35447–55. Epub 2011/08/25. doi: 10.1074/jbc.M111.274936 21862582

57. Bueter CL, Lee CK, Wang JP, Ostroff GR, Specht CA, Levitz SM. Spectrum and mechanisms of inflammasome activation by chitosan. J Immunol. 2014;192(12):5943–51. Epub 2014/05/16. doi: 10.4049/jimmunol.1301695 24829412

58. Singh A, Panting RJ, Varma A, Saijo T, Waldron KJ, Jong A, et al. Factors required for activation of urease as a virulence determinant in Cryptococcus neoformans. MBio. 2013;4(3):e00220–13. Epub 2013/05/09. doi: 10.1128/mBio.00220-13 23653445

59. Tangen KL, Jung WH, Sham AP, Lian T, Kronstad JW. The iron- and cAMP-regulated gene SIT1 influences ferrioxamine B utilization, melanization and cell wall structure in Cryptococcus neoformans. Microbiology. 2007;153(Pt 1):29–41. Epub 2006/12/23. doi: 10.1099/mic.0.2006/000927-0

60. Desalermos A, Tan X, Rajamuthiah R, Arvanitis M, Wang Y, Li D, et al. A Multi-Host Approach for the Systematic Analysis of Virulence Factors in Cryptococcus neoformans. J Infect Dis. 2014. Epub 2014/08/13. doi: 10.1093/infdis/jiu441

61. Fu J, Hettler E, Wickes BL. Split marker transformation increases homologous integration frequency in Cryptococcus neoformans. Fungal Genet Biol. 2006;43(3):200–12. Epub 2006/02/25. doi: 10.1016/j.fgb.2005.09.007 16497523

62. Toffaletti DL, Rude TH, Johnston SA, Durack DT, Perfect JR. Gene transfer in Cryptococcus neoformans by use of biolistic delivery of DNA. J Bacteriol. 1993;175(5):1405–11. Epub 1993/03/01. PMCID: 193227. 8444802

63. Skowyra ML, Doering TL. RNA interference in Cryptococcus neoformans. Methods Mol Biol. 2012;845:165–86. Epub 2012/02/14. doi: 10.1007/978-1-61779-539-8_11 22328374

64. Gerik KJ, Donlin MJ, Soto CE, Banks AM, Banks IR, Maligie MA, et al. Cell wall integrity is dependent on the PKC1 signal transduction pathway in Cryptococcus neoformans. Mol Microbiol. 2005;58(2):393–408. Epub 2005/10/01. doi: 10.1111/j.1365-2958.2005.04843.x 16194228

65. Feldmesser M, Rivera J, Kress Y, Kozel TR, Casadevall A. Antibody interactions with the capsule of Cryptococcus neoformans. Infect Immun. 2000;68(6):3642–50. Epub 2000/05/19. PMCID: 97654. 10816523