#PAGE_PARAMS# #ADS_HEAD_SCRIPTS# #MICRODATA#

Transcription Factor SomA Is Required for Adhesion, Development and Virulence of the Human Pathogen


Invasive fungal infections affecting immunocompromised patients are emerging worldwide. Among various human fungal pathogens, Aspergillus fumigatus is one of the most common molds causing severe invasive aspergillosis in immunocompromised patients. The conidia, which can evade from innate immunity and adhere to epithelial cells of alveoli in human lungs will start to germinate and cause the disease. Currently, the understanding of the molecular mechanisms of adherence of fungal cells to hosts is scarce. The transcription factor Flo8 controls adhesion to biotic or abiotic surfaces and morphological development in baker’s yeast. Flo8 homologues in the dimorphic human pathogenic yeast Candida albicans or the filamentous plant pathogen Magnaporthe oryzae are required for development and virulence. We found in this study that the Flo8 homologue SomA of A. fumigatus is required for adhesion and conidiation. Two independent invasive aspergillosis assays using chicken eggs or mouse demonstrated that deletion of the corresponding gene resulted in attenuated virulence. SomA represents an important fungal transcription factor at the interface between adherence, asexual spore formation and pathogenicity in an important opportunistic human pathogen.


Vyšlo v časopise: Transcription Factor SomA Is Required for Adhesion, Development and Virulence of the Human Pathogen. PLoS Pathog 11(11): e32767. doi:10.1371/journal.ppat.1005205
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1005205

Souhrn

Invasive fungal infections affecting immunocompromised patients are emerging worldwide. Among various human fungal pathogens, Aspergillus fumigatus is one of the most common molds causing severe invasive aspergillosis in immunocompromised patients. The conidia, which can evade from innate immunity and adhere to epithelial cells of alveoli in human lungs will start to germinate and cause the disease. Currently, the understanding of the molecular mechanisms of adherence of fungal cells to hosts is scarce. The transcription factor Flo8 controls adhesion to biotic or abiotic surfaces and morphological development in baker’s yeast. Flo8 homologues in the dimorphic human pathogenic yeast Candida albicans or the filamentous plant pathogen Magnaporthe oryzae are required for development and virulence. We found in this study that the Flo8 homologue SomA of A. fumigatus is required for adhesion and conidiation. Two independent invasive aspergillosis assays using chicken eggs or mouse demonstrated that deletion of the corresponding gene resulted in attenuated virulence. SomA represents an important fungal transcription factor at the interface between adherence, asexual spore formation and pathogenicity in an important opportunistic human pathogen.


Zdroje

1. de Groot PW, Bader O, de Boer AD, Weig M, Chauhan N. Adhesins in human fungal pathogens: glue with plenty of stick. Eukaryot Cell. 2013;12(4):470–81. Epub 2013/02/08. doi: 10.1128/EC.00364-12 23397570

2. Brückner S, Mösch HU. Choosing the right lifestyle: adhesion and development in Saccharomyces cerevisiae. FEMS Microbiol Rev. 2012;36(1):25–58. Epub 2011/05/20. doi: 10.1111/j.1574-6976.2011.00275.x 21521246

3. Thau N, Monod M, Crestani B, Rolland C, Tronchin G, Latgé JP, et al. rodletless mutants of Aspergillus fumigatus. Infect Immun. 1994;62(10):4380–8. Epub 1994/10/01. 7927699

4. Sheppard DC. Molecular mechanism of Aspergillus fumigatus adherence to host constituents. Curr Opin Microbiol. 2011;14(4):375–9. Epub 2011/07/23. doi: 10.1016/j.mib.2011.07.006 21784698

5. Aimanianda V, Bayry J, Bozza S, Kniemeyer O, Perruccio K, Elluru SR, et al. Surface hydrophobin prevents immune recognition of airborne fungal spores. Nature. 2009;460(7259):1117–21. doi: 10.1038/nature08264 19713928

6. Mayer FL, Wilson D, Hube B. Candida albicans pathogenicity mechanisms. Virulence. 2013;4(2):119–28. Epub 2013/01/09. doi: 10.4161/viru.22913 23302789

7. Li L, Wright SJ, Krystofova S, Park G, Borkovich KA. Heterotrimeric G protein signaling in filamentous fungi. Annu Rev Microbiol. 2007;61:423–52. Epub 2007/05/18. 17506673

8. Mitchell TK, Dean RA. The cAMP-dependent protein kinase catalytic subunit is required for appressorium formation and pathogenesis by the rice blast pathogen Magnaporthe grisea. Plant Cell. 1995;7(11):1869–78. Epub 1995/11/01. 8535140

9. Dürrenberger F, Wong K, Kronstad JW. Identification of a cAMP-dependent protein kinase catalytic subunit required for virulence and morphogenesis in Ustilago maydis. Proc Natl Acad Sci U S A. 1998;95(10):5684–9. Epub 1998/05/20. 9576944

10. Kozubowski L, Lee SC, Heitman J. Signalling pathways in the pathogenesis of Cryptococcus. Cell Microbiol. 2009;11(3):370–80. Epub 2008/12/19. doi: 10.1111/j.1462-5822.2008.01273.x 19170685

11. Hogan DA, Sundstrom P. The Ras/cAMP/PKA signaling pathway and virulence in Candida albicans. Future Microbiol. 2009;4(10):1263–70. Epub 2009/12/10. doi: 10.2217/fmb.09.106 19995187

12. Ramanujam R, Naqvi NI. PdeH, a high-affinity cAMP phosphodiesterase, is a key regulator of asexual and pathogenic differentiation in Magnaporthe oryzae. PLoS Pathog. 2010;6(5):e1000897. Epub 2010/05/14. doi: 10.1371/journal.ppat.1000897 20463817

13. Liebmann B, Gattung S, Jahn B, Brakhage AA. cAMP signaling in Aspergillus fumigatus is involved in the regulation of the virulence gene pksP and in defense against killing by macrophages. Mol Genet Genomics. 2003;269(3):420–35. Epub 2003/05/07. 12734751

14. Zhao W, Panepinto JC, Fortwendel JR, Fox L, Oliver BG, Askew DS, et al. Deletion of the regulatory subunit of protein kinase A in Aspergillus fumigatus alters morphology, sensitivity to oxidative damage, and virulence. Infect Immun. 2006;74(8):4865–74. Epub 2006/07/25. 16861675

15. Fuller KK, Richie DL, Feng X, Krishnan K, Stephens TJ, Wikenheiser-Brokamp KA, et al. Divergent Protein Kinase A isoforms co-ordinately regulate conidial germination, carbohydrate metabolism and virulence in Aspergillus fumigatus. Mol Microbiol. 2011;79(4):1045–62. Epub 2011/01/06. doi: 10.1111/j.1365-2958.2010.07509.x 21210869

16. Liebmann B, Müller M, Braun A, Brakhage AA. The cyclic AMP-dependent protein kinase a network regulates development and virulence in Aspergillus fumigatus. Infect Immun. 2004;72(9):5193–203. Epub 2004/08/24. 15322014

17. 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. Epub 2010/03/12. doi: 10.1086/651263 20218877

18. Pappas PG, Alexander BD, Andes DR, Hadley S, Kauffman CA, Freifeld A, et al. Invasive fungal infections among organ transplant recipients: results of the Transplant-Associated Infection Surveillance Network (TRANSNET). Clin Infect Dis. 2010;50(8):1101–11. Epub 2010/03/12. doi: 10.1086/651262 20218876

19. Tekaia F, Latgé JP. Aspergillus fumigatus: saprophyte or pathogen? Curr Opin Microbiol. 2005;8(4):385–92. Epub 2005/07/16. 16019255

20. Cao F, Lane S, Raniga PP, Lu Y, Zhou Z, Ramon K, et al. The Flo8 transcription factor is essential for hyphal development and virulence in Candida albicans. Mol Biol Cell. 2006;17(1):295–307. Epub 2005/11/02. 16267276

21. Shapiro RS, Ryan O, Boone C, Cowen LE. Regulatory circuitry governing morphogenesis in Saccharomyces cerevisiae and Candida albicans. Cell Cycle. 2012;11(23):4294–5. Epub 2012/10/24. doi: 10.4161/cc.22608 23095675

22. Ryan O, Shapiro RS, Kurat CF, Mayhew D, Baryshnikova A, Chin B, et al. Global gene deletion analysis exploring yeast filamentous growth. Science. 2012;337(6100):1353–6. Epub 2012/09/18. doi: 10.1126/science.1224339 22984072

23. Yan X, Li Y, Yue X, Wang C, Que Y, Kong D, et al. Two novel transcriptional regulators are essential for infection-related morphogenesis and pathogenicity of the rice blast fungus Magnaporthe oryzae. PLoS Pathog. 2011;7(12):e1002385. Epub 2011/12/01. doi: 10.1371/journal.ppat.1002385 22144889

24. Bayram Ö, Braus GH. Coordination of secondary metabolism and development in fungi: the velvet family of regulatory proteins. FEMS Microbiol Rev. 2012;36(1):1–24. Epub 2011/06/11. doi: 10.1111/j.1574-6976.2011.00285.x 21658084

25. Yu JH. Regulation of Development in Aspergillus nidulans and Aspergillus fumigatus. Mycobiology. 2010;38(4):229–37. Epub 2010/12/31. doi: 10.4489/MYCO.2010.38.4.229 23956662

26. Mah JH, Yu JH. Upstream and downstream regulation of asexual development in Aspergillus fumigatus. Eukaryot Cell. 2006;5(10):1585–95. Epub 2006/10/13. 17030990

27. Adams TH, Wieser JK, Yu JH. Asexual sporulation in Aspergillus nidulans. Microbiol Mol Biol Rev. 1998;62(1):35–54. Epub 1998/04/08. 9529886

28. Gravelat FN, Ejzykowicz DE, Chiang LY, Chabot JC, Urb M, Macdonald KD, et al. Aspergillus fumigatus MedA governs adherence, host cell interactions and virulence. Cell Microbiol. 2010;12(4):473–88. Epub 2009/11/04. doi: 10.1111/j.1462-5822.2009.01408.x 19889083

29. Sheppard DC, Doedt T, Chiang LY, Kim HS, Chen D, Nierman WC, et al. The Aspergillus fumigatus StuA protein governs the up-regulation of a discrete transcriptional program during the acquisition of developmental competence. Mol Biol Cell. 2005;16(12):5866–79. Epub 2005/10/05. 16207816

30. Gravelat FN, Beauvais A, Liu H, Lee MJ, Snarr BD, Chen D, et al. Aspergillus galactosaminogalactan mediates adherence to host constituents and conceals hyphal β-glucan from the immune system. PLoS Pathog. 2013;9(8):e1003575. Epub 2013/08/22. doi: 10.1371/journal.ppat.1003575 23990787

31. Liu H, Styles CA, Fink GR. Saccharomyces cerevisiae S288C has a mutation in FLO8, a gene required for filamentous growth. Genetics. 1996;144(3):967–78. Epub 1996/11/01. 8913742

32. Fichtner L, Schulze F, Braus GH. Differential Flo8p-dependent regulation of FLO1 and FLO11 for cell-cell and cell-substrate adherence of S. cerevisiae S288c. Mol Microbiol. 2007;66(5):1276–89. Epub 2007/11/16. 18001350

33. Bester MC, Pretorius IS, Bauer FF. The regulation of Saccharomyces cerevisiae FLO gene expression and Ca2+ -dependent flocculation by Flo8p and Mss11p. Curr Genet. 2006;49(6):375–83. Epub 2006/03/25. 16568252

34. Rupp S, Summers E, Lo HJ, Madhani H, Fink G. MAP kinase and cAMP filamentation signaling pathways converge on the unusually large promoter of the yeast FLO11 gene. EMBO J. 1999;18(5):1257–69. Epub 1999/03/04. 10064592

35. Twumasi-Boateng K, Yu Y, Chen D, Gravelat FN, Nierman WC, Sheppard DC. Transcriptional profiling identifies a role for BrlA in the response to nitrogen depletion and for StuA in the regulation of secondary metabolite clusters in Aspergillus fumigatus. Eukaryot Cell. 2009;8(1):104–15. Epub 2008/11/21. doi: 10.1128/EC.00265-08 19028996

36. Helmschrott C, Sasse A, Samantaray S, Krappmann S, Wagener J. Upgrading fungal gene expression on demand: improved systems for doxycycline-dependent silencing in Aspergillus fumigatus. Appl Environ Microbiol. 2013;79(5):1751–4. Epub 2012/12/28. doi: 10.1128/AEM.03626-12 23275515

37. Park HS, Bayram Ö, Braus GH, Kim SC, Yu JH. Characterization of the velvet regulators in Aspergillus fumigatus. Mol Microbiol. 2012;86(4):937–53. Epub 2012/09/14. doi: 10.1111/mmi.12032 22970834

38. Ahmed YL, Gerke J, Park HS, Bayram Ö, Neumann P, Ni M, et al. The velvet family of fungal regulators contains a DNA-binding domain structurally similar to NF-κB. PLoS Biol. 2013;11(12):e1001750. Epub 2013/12/31. doi: 10.1371/journal.pbio.1001750 24391470

39. Chaudhuri R, Ansari FA, Raghunandanan MV, Ramachandran S. FungalRV: Adhesin prediction and immunoinformatics portal for human fungal pathogens. BMC Genomics. 2011;12(1):192–205. Epub 2011/04/19.

40. Jacobsen ID, Große K, Slesiona S, Hube B, Berndt A, Brock M. Embryonated eggs as an alternative infection model to investigate Aspergillus fumigatus virulence. Infect Immun. 2010;78(7):2995–3006. Epub 2010/04/26. doi: 10.1128/IAI.00268-10 20421382

41. Kupfahl C, Heinekamp T, Geginat G, Ruppert T, Hartl A, Hof H, et al. Deletion of the gliP gene of Aspergillus fumigatus results in loss of gliotoxin production but has no effect on virulence of the fungus in a low-dose mouse infection model. Mol Microbiol. 2006;62(1):292–302. Epub 2006/08/31. 16956378

42. Ben-Ami R, Lewis RE, Leventakos K, Latge JP, Kontoyiannis DP. Cutaneous model of invasive aspergillosis. Antimicrob Agents Chemother. 2010;54(5):1848–54. Epub 2010/02/11. doi: 10.1128/AAC.01504-09 20145078

43. Wagener J, Echtenacher B, Rohde M, Kotz A, Krappmann S, Heesemann J, et al. The putative alpha-1,2-mannosyltransferase AfMnt1 of the opportunistic fungal pathogen Aspergillus fumigatus is required for cell wall stability and full virulence. Eukaryot Cell. 2008;7(10):1661–73. Epub 2008/08/19. doi: 10.1128/EC.00221-08 18708564

44. Lee BY, Han SY, Choi HG, Kim JH, Han KH, Han DM. Screening of growth- or development-related genes by using genomic library with inducible promoter in Aspergillus nidulans. J Microbiol. 2005;43(6):523–8. Epub 2006/01/18. 16410769

45. Li F, Palecek SP. Identification of Candida albicans genes that induce Saccharomyces cerevisiae cell adhesion and morphogenesis. Biotechnol Prog. 2005;21(6):1601–9. Epub 2005/12/03. 16321041

46. Tran VT, Braus-Stromeyer SA, Kusch H, Reusche M, Kaever A, Kuhn A, et al. Verticillium transcription activator of adhesion Vta2 suppresses microsclerotia formation and is required for systemic infection of plant roots. New Phytol. 2014;202(2):565–81. Epub 2014/01/17. doi: 10.1111/nph.12671 24433459

47. Pan X, Heitman J. Protein kinase A operates a molecular switch that governs yeast pseudohyphal differentiation. Mol Cell Biol. 2002;22(12):3981–93. Epub 2002/05/25. 12024012

48. Pan X, Heitman J. Cyclic AMP-dependent protein kinase regulates pseudohyphal differentiation in Saccharomyces cerevisiae. Mol Cell Biol. 1999;19(7):4874–87. Epub 1999/06/22. 10373537

49. Kim TS, Kim HY, Yoon JH, Kang HS. Recruitment of the Swi/Snf complex by Ste12-Tec1 promotes Flo8-Mss11-mediated activation of STA1 expression. Mol Cell Biol. 2004;24(21):9542–56. Epub 2004/10/16. 15485921

50. Govender P, Domingo JL, Bester MC, Pretorius IS, Bauer FF. Controlled expression of the dominant flocculation genes FLO1, FLO5, and FLO11 in Saccharomyces cerevisiae. Appl Environ Microbiol. 2008;74(19):6041–52. Epub 2008/08/15. doi: 10.1128/AEM.00394-08 18708514

51. Jurata LW, Kenny DA, Gill GN. Nuclear LIM interactor, a rhombotin and LIM homeodomain interacting protein, is expressed early in neuronal development. Proc Natl Acad Sci U S A. 1996;93(21):11693–8. Epub 1996/10/15. 8876198

52. van Meyel DJ, Thomas JB, Agulnick AD. Ssdp proteins bind to LIM-interacting co-factors and regulate the activity of LIM-homeodomain protein complexes in vivo. Development. 2003;130(9):1915–25. Epub 2003/03/19. 12642495

53. Su C, Li Y, Lu Y, Chen J. Mss11, a transcriptional activator, is required for hyphal development in Candida albicans. Eukaryot Cell. 2009;8(11):1780–91. Epub 2009/09/04. doi: 10.1128/EC.00190-09 19734367

54. Radivojac P, Vacic V, Haynes C, Cocklin RR, Mohan A, Heyen JW, et al. Identification, analysis, and prediction of protein ubiquitination sites. Proteins. 2010;78(2):365–80. Epub 2009/09/02. doi: 10.1002/prot.22555 19722269

55. Tao L, Yu JH. AbaA and WetA govern distinct stages of Aspergillus fumigatus development. Microbiology. 2011;157(Pt 2):313–26. Epub 2010/10/21. doi: 10.1099/mic.0.044271-0 20966095

56. Xiao P, Shin KS, Wang T, Yu JH. Aspergillus fumigatus flbB encodes two basic leucine zipper domain (bZIP) proteins required for proper asexual development and gliotoxin production. Eukaryot Cell. 2010;9(11):1711–23. Epub 2010/09/17. doi: 10.1128/EC.00198-10 20852021

57. Garzia A, Etxebeste O, Herrero-Garcia E, Fischer R, Espeso EA, Ugalde U. Aspergillus nidulans FlbE is an upstream developmental activator of conidiation functionally associated with the putative transcription factor FlbB. Mol Microbiol. 2009;71(1):172–84. Epub 2008/11/05. doi: 10.1111/j.1365-2958.2008.06520.x 19007409

58. Garzia A, Etxebeste O, Herrero-Garcia E, Ugalde U, Espeso EA. The concerted action of bZip and cMyb transcription factors FlbB and FlbD induces brlA expression and asexual development in Aspergillus nidulans. Mol Microbiol. 2010;75(5):1314–24. Epub 2010/02/01. doi: 10.1111/j.1365-2958.2010.07063.x 20132447

59. Krijgsheld P, Bleichrodt R, van Veluw GJ, Wang F, Müller WH, Dijksterhuis J, et al. Development in Aspergillus. Stud Mycol. 2013;74(1):1–29. Epub 2013/03/02. doi: 10.3114/sim0006 23450714

60. Loussert C, Schmitt C, Prevost MC, Balloy V, Fadel E, Philippe B, et al. In vivo biofilm composition of Aspergillus fumigatus. Cell Microbiol. 2010;12(3):405–10. Epub 2009/11/04. doi: 10.1111/j.1462-5822.2009.01409.x 19889082

61. Dutton JR, Johns S, Miller BL. StuAp is a sequence-specific transcription factor that regulates developmental complexity in Aspergillus nidulans. EMBO J. 1997;16(18):5710–21. Epub 1997/10/06. 9312029

62. Macheleidt J, Scherlach K, Neuwirth T, Schmidt-Heck W, Straßburger M, Spraker J, et al. Transcriptome analysis of cyclic AMP-dependent protein kinase A-regulated genes reveals the production of the novel natural compound fumipyrrole by Aspergillus fumigatus. Mol Microbiol. 2015;96(1):148–62. Epub 2015/03/11. doi: 10.1111/mmi.12926 25582336

63. Gimeno CJ, Fink GR. Induction of pseudohyphal growth by overexpression of PHD1, a Saccharomyces cerevisiae gene related to transcriptional regulators of fungal development. Mol Cell Biol. 1994;14(3):2100–12. Epub 1994/03/01. 8114741

64. Gavrias V, Andrianopoulos A, Gimeno CJ, Timberlake WE. Saccharomyces cerevisiae TEC1 is required for pseudohyphal growth. Mol Microbiol. 1996;19(6):1255–63. Epub 1996/03/01. 8730867

65. Linder MB, Szilvay GR, Nakari-Setälä T, Penttilä ME. Hydrophobins: the protein-amphiphiles of filamentous fungi. FEMS Microbiol Rev. 2005;29(5):877–96. Epub 2005/02/21. 16219510

66. Kim S, Ahn IP, Rho HS, Lee YH. MHP1, a Magnaporthe grisea hydrophobin gene, is required for fungal development and plant colonization. Mol Microbiol. 2005;57(5):1224–37. Epub 2005/08/17. 16101997

67. Talbot NJ, Ebbole DJ, Hamer JE. Identification and characterization of MPG1, a gene involved in pathogenicity from the rice blast fungus Magnaporthe grisea. Plant Cell. 1993;5(11):1575–90. Epub 1993/11/01. 8312740

68. Gauthier GM, Keller NP. Crossover fungal pathogens: the biology and pathogenesis of fungi capable of crossing kingdoms to infect plants and humans. Fungal Genet Biol. 2013;61:146–57. Epub 2013/09/08. doi: 10.1016/j.fgb.2013.08.016 24021881

69. Sexton AC, Howlett BJ. Parallels in fungal pathogenesis on plant and animal hosts. Eukaryot Cell. 2006;5(12):1941–9. Epub 2006/10/13. 17041185

70. Bayry J, Aimanianda V, Guijarro JI, Sunde M, Latgé JP. Hydrophobins—unique fungal proteins. PLoS Pathog. 2012;8(5):e1002700. Epub 2012/05/31. doi: 10.1371/journal.ppat.1002700 22693445

71. Haas H, Eisendle M, Turgeon BG. Siderophores in fungal physiology and virulence. Annu Rev Phytopathol. 2008;46:149–87. Epub 2008/08/06. doi: 10.1146/annurev.phyto.45.062806.094338 18680426

72. Krappmann S, Sasse C, Braus GH. Gene targeting in Aspergillus fumigatus by homologous recombination is facilitated in a nonhomologous end- joining-deficient genetic background. Eukaryot Cell. 2006;5(1):212–5. Epub 2006/01/10. 16400185

73. Käfer E. Meiotic and mitotic recombination in Aspergillus and its chromosomal aberrations. Adv Genet. 1977;19:33–131. Epub 1977/01/01. 327767

74. Krappmann S, Bayram Ö, Braus GH. Deletion and allelic exchange of the Aspergillus fumigatus veA locus via a novel recyclable marker module. Eukaryot Cell. 2005;4(7):1298–307. Epub 2005/07/09. 16002655

75. Braus GH, Grundmann O, Brückner S, Mösch HU. Amino acid starvation and Gcn4p regulate adhesive growth and FLO11 gene expression in Saccharomyces cerevisiae. Mol Biol Cell. 2003;14(10):4272–84. Epub 2003/06/27. 14517335

76. Guo B, Styles CA, Feng Q, Fink GR. A Saccharomyces gene family involved in invasive growth, cell-cell adhesion, and mating. Proc Natl Acad Sci U S A. 2000;97(22):12158–63. Epub 2000/10/12. 11027318

77. Sambrook J, Fritsch E, Maniatis T. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press: New York; 1989.

78. Lee B, Taylor J. Isolation of DNA from fungal mycelia and single spores. Innis M, Gelfand D, Sninsky J, White T, editors. Academic Press Inc: San Diego; 1990. 282–7 p.

79. Hartmann T, Dümig M, Jaber BM, Szewczyk E, Olbermann P, Morschhäuser J, et al. Validation of a self-excising marker in the human pathogen Aspergillus fumigatus by employing the β-rec/six site-specific recombination system. Appl Environ Microbiol. 2010;76(18):6313–7. Epub 2010/07/23. doi: 10.1128/AEM.00882-10 20656854

80. Yelton MM, Hamer JE, Timberlake WE. Transformation of Aspergillus nidulans by using a trpC plasmid. Proc Natl Acad Sci U S A. 1984;81(5):1470–4. Epub 1984/03/01. 6324193

81. Southern EM. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975;98(3):503–17. Epub 1975/11/05. 1195397

82. Arnaud MB, Cerqueira GC, Inglis DO, Skrzypek MS, Binkley J, Chibucos MC, et al. The Aspergillus Genome Database (AspGD): recent developments in comprehensive multispecies curation, comparative genomics and community resources. Nucleic Acids Res. 2012;40(Database issue):D653–9. Epub 2011/11/12. doi: 10.1093/nar/gkr875 22080559

83. Ito H, Fukuda Y, Murata K, Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983;153(1):163–8. Epub 1983/01/01. 6336730

84. Rose M, Botstein D. Construction and use of gene fusions to lacZ (β-galactosidase) that are expressed in yeast. Methods Enzymol. 1983;101:167–80. Epub 1983/01/01. 6310320

85. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72:248–54. Epub 1976/05/07. 942051

86. Shevchenko A, Wilm M, Vorm O, Mann M. Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels. Anal Chem. 1996;68(5):850–8. Epub 1996/03/01. 8779443

87. von Zeska Kress MR, Harting R, Bayram Ö, Christmann M, Irmer H, Valerius O, et al. The COP9 signalosome counteracts the accumulation of cullin SCF ubiquitin E3 RING ligases during fungal development. Mol Microbiol. 2012;83(6):1162–77. Epub 2012/02/22. doi: 10.1111/j.1365-2958.2012.07999.x 22329854

88. Tesfaigzi J, Smith-Harrison W, Carlson DM. A simple method for reusing western blots on PVDF membranes. Biotechniques. 1994;17(2):268–9. Epub 1994/08/01. 7980922

89. Harting R, Bayram Ö, Laubinger K, Valerius O, Braus GH. Interplay of the fungal sumoylation network for control of multicellular development. Mol Microbiol. 2013;90(5):1125–45. Epub 2013/11/06. doi: 10.1111/mmi.12421 24279728

90. Cox J, Mann M. MaxQuant enables high peptide identification rates, individualized p.p.b.-range mass accuracies and proteome-wide protein quantification. Nat Biotechnol. 2008;26(12):1367–72. Epub 2008/11/30. doi: 10.1038/nbt.1511 19029910

91. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-ΔΔC(T)) Method. Methods. 2001;25(4):402–8. Epub 2002/02/16. 11846609

92. Fontaine T, Delangle A, Simenel C, Coddeville B, van Vliet SJ, van Kooyk Y, et al. Galactosaminogalactan, a new immunosuppressive polysaccharide of Aspergillus fumigatus. PLoS Pathog. 2011;7(11):e1002372. Epub 2011/11/22. doi: 10.1371/journal.ppat.1002372 22102815

93. Mumberg D, Müller R, Funk M. Regulatable promoters of Saccharomyces cerevisiae: comparison of transcriptional activity and their use for heterologous expression. Nucleic Acids Res. 1994;22(25):5767–8. Epub 1994/12/25. 7838736

Štítky
Hygiena a epidemiológia Infekčné lekárstvo Laboratórium

Článok vyšiel v časopise

PLOS Pathogens


2015 Číslo 11
Najčítanejšie tento týždeň
Najčítanejšie v tomto čísle
Kurzy

Zvýšte si kvalifikáciu online z pohodlia domova

Získaná hemofilie - Povědomí o nemoci a její diagnostika
nový kurz

Eozinofilní granulomatóza s polyangiitidou
Autori: doc. MUDr. Martina Doubková, Ph.D.

Všetky kurzy
Prihlásenie
Zabudnuté heslo

Zadajte e-mailovú adresu, s ktorou ste vytvárali účet. Budú Vám na ňu zasielané informácie k nastaveniu nového hesla.

Prihlásenie

Nemáte účet?  Registrujte sa

#ADS_BOTTOM_SCRIPTS#