Skip to main content

Fungal Cell Wall Analysis

  • Chapter
  • First Online:
Laboratory Protocols in Fungal Biology

Part of the book series: Fungal Biology ((FUNGBIO))

Abstract

Fungal cell wall is a rigid structure mainly composed of polysaccharides (up to 90 %) and glycoproteins. It is essential for survival of the fungal cells, because it protects them against bursting caused by internal turgor pressure and against mechanical injury. Because of its absence in mammalian cells, it is an attractive target for antifungal agents. Thus, for various reasons, it might be important to know how the cell wall is synthesized, and how to analyze its composition. We provide here information about in vitro analysis of the biosynthetic activities of the main fungal wall and describe some methods for rapid analysis of cell wall composition by using specific enzymatic degradations. We also describe some additional methods that can be occasionally used to analyze fungal wall properties or composition. These methods provide powerful tools to evaluate changes in fungal cell walls and will be useful for screening new compounds for antifungal activity that might cause inhibition of cell wall biosynthesis and/or alter the structure of the fungal cell wall.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 189.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 249.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 329.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Latge JP (2007) The cell wall: a carbohydrate armour for the fungal cell. Mol Microbiol 66:279–290

    Article  PubMed  CAS  Google Scholar 

  2. Cabib E, Bowers B, Sburlati A, Silverman SJ (1988) Fungal cell wall synthesis: the construction of a biological structure. Microbiol Sci 5:370–375

    PubMed  CAS  Google Scholar 

  3. Lesage G, Bussey H (2006) Cell wall assembly in Saccharomyces cerevisiae. Microbiol Mol Biol Rev 70:317–343

    Article  PubMed  CAS  Google Scholar 

  4. Cabib E, Kang MS (1987) Fungal 1,3-b-glucan synthase. Methods Enzymol 138:637–642

    Article  PubMed  CAS  Google Scholar 

  5. Kapteyn JC, Ram AF, Groos EM, Kollar R, Montijn RC, Van Den Ende H et al (1997) Altered extent of cross-linking of b1,6-glucosylated mannoproteins to chitin in Saccharomyces cerevisiae mutants with reduced cell wall b1,3-glucan content. J Bacteriol 179:6279–6284

    PubMed  CAS  Google Scholar 

  6. Kollar R, Reinhold BB, Petráková E, Yeh HJ, Ashwell G, Drgonová J et al (1997) Architecture of the yeast cell wall. b(1-6)-glucan interconnects mannoprotein, b(1-3)-glucan, and chitin. J Biol Chem 272:17762–17775

    Article  PubMed  CAS  Google Scholar 

  7. Kollar R, Petrakova E, Ashwell G, Robbins PW, Cabib E (1995) Architecture of the yeast cell wall. The linkage between chitin and b(1-3)-glucan. J Biol Chem 270:1170–1178

    Article  PubMed  CAS  Google Scholar 

  8. Cabib E, Blanco N, Grau C, Rodriguez-Pena JM, Arroyo J (2007) Crh1p and Crh2p are required for the cross-linking of chitin to b(1-6)glucan in the Saccharomyces cerevisiae cell wall. Mol Microbiol 63:921–935

    Article  PubMed  CAS  Google Scholar 

  9. Cabib E, Farkas V, Kosik O, Blanco N, Arroyo J, McPhie P (2008) Assembly of the yeast cell wall. Crh1p and Crh2p act as transglycosylases in vivo and in vitro. J Biol Chem 283:29859–29872

    Article  PubMed  CAS  Google Scholar 

  10. Kapteyn JC, Montijn RC, Vink E, de la Cruz J, Llobell A, Douwes JE, Shimoi H, Lipke PN, Klis FM (1996) Retention of Saccharomyces cerevisiae cell wall proteins through a phosphodiester-linked b-1,3-/b-1,6-glucan heteropolymer. Glycobiology 6:337–345

    Article  PubMed  CAS  Google Scholar 

  11. Klis FM, Boorsma A, De Groot PW (2006) Cell wall construction in Saccharomyces cerevisiae. Yeast 23:185–202

    Article  PubMed  CAS  Google Scholar 

  12. Douglas CM, Marrinan JA, Li W, Kurtz MB (1994) A Saccharomyces cerevisiae mutant with echinocandin-resistant 1,3-b-D-glucan synthase. J Bacteriol 176:5686–5696

    PubMed  CAS  Google Scholar 

  13. Mazur P, Morin N, Baginsky W, El-Sherbeini M, Clemas JA, Nielsen JB et al (1995) Differential expression and function of two homologous subunits of yeast 1,3-b-D-glucan synthase. Mol Cell Biol 15:5671–5681

    PubMed  CAS  Google Scholar 

  14. Pérez P, Ribas JC (2004) Cell wall analysis. Methods 33:245–251

    Article  PubMed  CAS  Google Scholar 

  15. Arellano M, Duran A, Perez P (1996) Rho1 GTPase activates the (1-3)b-D-glucan synthase and is involved in Schizosaccharomyces pombe morphogenesis. EMBO J 15:4584–4591

    PubMed  CAS  Google Scholar 

  16. Qadota H, Python CP, Inoue SB, Arisawa M, Anraku Y, Zheng Y et al (1996) Identification of yeast Rho1p GTPase as a regulatory subunit of 1,3-b-glucan synthase. Science 272:279–281

    Article  PubMed  CAS  Google Scholar 

  17. Katayama S, Hirata D, Arellano M, Pérez P, Toda T (1999) Fission yeast a-glucan synthase Mok1 requires the actin cytoskeleton to localize the sites of growth and plays an essential role in cell morphogenesis downstream of protein kinase C function. J Cell Biol 144:1173–1186

    Article  PubMed  CAS  Google Scholar 

  18. Hoschstenbach F, Klis FM, Van den Ende H, Van Donselaar E, Peters PJ et al (1998) Identification of a putative alpha-glucan synthase essential for cell wall construction and morphogenesis in fission yeast. Proc Natl Acad Sci USA 95:9161–9166

    Article  Google Scholar 

  19. Grun CH, Hochstenbach F, Humbel BM, Verkleij AJ, Sietsma JH, Klis FM et al (2005) The structure of cell wall alpha-glucan from fission yeast. Glycobiology 15:245–257

    Article  PubMed  Google Scholar 

  20. Vos A, Dekker N, Distel B, Leunissen JA, Hochstenbach F (2007) Role of the synthase domain of Ags1p in cell wall alpha-glucan biosynthesis in fission yeast. J Biol Chem 282:18969–18979

    Article  PubMed  CAS  Google Scholar 

  21. Cabib E, Roh DH, Schmidt M, Crotti LB, Varma A (2001) The yeast cell wall and septum as paradigms of cell growth and morphogenesis. J Biol Chem 276:19679–19682

    Article  PubMed  CAS  Google Scholar 

  22. Roncero C (2002) The genetic complexity of chitin synthesis in fungi. Curr Genet 41:367–378

    Article  PubMed  CAS  Google Scholar 

  23. Lenardon MD, Munro CA, Gow NA (2010) Chitin synthesis and fungal pathogenesis. Curr Opin Microbiol 13:416–423

    Article  PubMed  CAS  Google Scholar 

  24. Bowman SM, Free SJ (2006) The structure and ­synthesis of the fungal cell wall. Bioessays 28:799–808

    Article  PubMed  Google Scholar 

  25. Leal JA, Prieto A, Bernabe M, Hawksworth DL (2010) An assessment of fungal wall heteromannans as a phylogenetically informative character in ascomycetes. FEMS Microbiol Rev 34:986–1014

    PubMed  CAS  Google Scholar 

  26. Shematek EM, Braatz JA, Cabib E (1980) Biosynthesis of yeast cell wall. I. Preparation and properties of b(1-3)glucan synthetase. J Biol Chem 255:888–894

    PubMed  CAS  Google Scholar 

  27. Shematek EM, Cabib E (1980) Biosynthesis of yeast cell wall. II. Regulation of b(1-3)glucan synthetase by ATP and GTP. J Biol Chem 255:895–902

    PubMed  CAS  Google Scholar 

  28. Ishiguro J, Saitou A, Durán A, Ribas JC (1997) cps1 +, a Schizosaccharomyces pombe gene homolog of Saccharomyces cerevisiae FKS genes whose mutation confers hypersensitivity to cyclosporin A and papulacandin B. J Bacteriol 179:7653–7662

    PubMed  CAS  Google Scholar 

  29. Martins IM, Cortés JCG, Muñoz J, Moreno MB, Ramos M, Clemente-Ramos JA et al (2011) Differential activities of three families of specific b(1,3)glucan synthase inhibitors in wild-type and resistant strains of fission yeast. J Biol Chem 286:3484–3496

    Article  PubMed  CAS  Google Scholar 

  30. Abe M, Nishida I, Minemura M, Qadota H, Seyama Y, Watanabe T et al (2001) Yeast 1,3-b-glucan synthase activity is inhibited by phytosphingosine localized to the endoplasmic reticulum. J Biol Chem 276:26923–26930

    Article  PubMed  CAS  Google Scholar 

  31. El-Sherbeini M, Clemas JA (1995) Nikkomycin Z supersensitivity of an echinocandin-resistant mutant of Saccharomyces cerevisiae. Antimicrob Agents Chemother 39:200–207

    Article  PubMed  CAS  Google Scholar 

  32. Inoue SB, Takewaki N, Takasuka T, Mio T, Adachi M, Fujii Y et al (1995) Characterization and gene cloning of 1,3-b-D-glucan synthase from Saccharomyces cerevisiae. Eur J Biochem 231:845–854

    Article  PubMed  CAS  Google Scholar 

  33. Kelly R, Register E, Hsu MJ, Kurtz M, Nielsen J (1996) Isolation of a gene involved in 1,3-b-glucan synthesis in Aspergillus nidulans and purification of the corresponding protein. J Bacteriol 178:4381–4391

    PubMed  CAS  Google Scholar 

  34. Mazur P, Baginsky W (1996) In vitro activity of 1,3-b-D-glucan synthase requires the GTP-binding protein Rho1. J Biol Chem 271:14604–14609

    Article  PubMed  CAS  Google Scholar 

  35. Mol PC, Park HM, Mullins JT, Cabib E (1994) A GTP-binding protein regulates the activity of (1,3)-b-glucan synthase, an enzyme directly involved in yeast cell wall morphogenesis. J Biol Chem 269:31267–31274

    PubMed  CAS  Google Scholar 

  36. Thompson JR, Douglas CM, Li W, Jue CK, Pramanik B, Yuan X et al (1999) A glucan synthase FKS1 homolog in Cryptococcus neoformans is single copy and encodes an essential function. J Bacteriol 181:444–453

    PubMed  CAS  Google Scholar 

  37. Wood RL, Miller TK, Wright A, McCarthy P, Taft CS, Pomponi S et al (1998) Characterization and optimization of in vitro assay conditions for (1,3)b-glucan synthase activity from Aspergillus fumigatus and Candida albicans for enzyme inhibition screening. J Antibiot (Tokyo) 51:665–675

    Article  CAS  Google Scholar 

  38. Sestak S, Farkas V (2001) In situ assays of fungal enzymes in cells permeabilized by osmotic shock. Anal Biochem 292:34–39

    Article  PubMed  CAS  Google Scholar 

  39. Shedletzky E, Unger C, Delmer DP (1997) A microtiter-based fluorescence assay for (1,3)-b-glucan synthases. Anal Biochem 249:88–93

    Article  PubMed  CAS  Google Scholar 

  40. Ribas JC, Díaz M, Durán A, Pérez P (1991) Isolation and characterization of Schizosaccharomyces pombe mutants defective in cell wall (1-3)b-D-glucan. J Bacteriol 173:3456–3462

    PubMed  CAS  Google Scholar 

  41. El-Sherbeini M, Clemas JA (1995) Cloning and characterization of GNS1: a Saccharomyces cerevisiae gene involved in synthesis of 1,3-b-glucan in vitro. J Bacteriol 177:3227–3234

    PubMed  CAS  Google Scholar 

  42. Kang MS, Cabib E (1986) Regulation of fungal cell wall growth: a guanine nucleotide-binding proteinaceous component required for activity of (1,3)-b-D-glucan synthase. Proc Natl Acad Sci U S A 83:5808–5812

    Article  PubMed  CAS  Google Scholar 

  43. Kondoh O, Tachibana Y, Ohya Y, Arisawa M, Watanabe T (1997) Cloning of the RHO1 gene from Candida albicans and its regulation of b-1,3-glucan synthesis. J Bacteriol 179:7734–7741

    PubMed  CAS  Google Scholar 

  44. Schimoler-O’Rourke R, Renault S, Mo W, Selitrennikoff CP (2003) Neurospora crassa FKS protein binds to the (1,3)b-glucan synthase substrate, UDP-glucose. Curr Microbiol 46:408–412

    Article  PubMed  CAS  Google Scholar 

  45. Crotti LB, Drgon T, Cabib E (2001) Yeast cell permeabilization by osmotic shock allows determination of enzymatic activities in situ. Anal Biochem 292:8–16

    Article  PubMed  CAS  Google Scholar 

  46. Aimanianda V, Clavaud C, Simenel C, Fontaine T, Delepierre M, Latge JP (2009) Cell wall b-(1,6)-­glucan of Saccharomyces cerevisiae: structural characterization and in situ synthesis. J Biol Chem 284:13401–13412

    Article  PubMed  CAS  Google Scholar 

  47. Frost DJ, Brandt K, Capobianco J, Goldman R (1994) Characterization of (1,3)-b-glucan synthase in Candida albicans: microsomal assay from the yeast or mycelial morphological forms and a permeabilized whole-cell assay. Microbiology 140:2239–2246

    Article  PubMed  CAS  Google Scholar 

  48. Choi WJ, Cabib E (1994) The use of divalent cations and pH for the determination of specific yeast chitin synthetases. Anal Biochem 219:368–372

    Article  PubMed  CAS  Google Scholar 

  49. Lucero HA, Kuranda MJ, Bulik DA (2002) A nonradioactive, high throughput assay for chitin synthase activity. Anal Biochem 305:97–105

    Article  PubMed  CAS  Google Scholar 

  50. Castro C, Ribas JC, Valdivieso MH, Varona R, del Rey F, Durán A (1995) Papulacandin B resistance in budding and fission yeasts: isolation and characterization of a gene involved in (1,3)b-D-glucan synthesis in Saccharomyces cerevisiae. J Bacteriol 177:5732–5739

    PubMed  CAS  Google Scholar 

  51. Roncero C, Valdivieso MH, Ribas JC, Duran A (1988) Isolation and characterization of Saccharomyces ­cerevisiae mutants resistant to Calcofluor white. J Bacteriol 170:1950–1954

    PubMed  CAS  Google Scholar 

  52. Algranati ID, Behrens N, Carminatti H, Cabib E (1966) Mannan synthetase from yeast. Methods Enzymol 8:411–416

    Article  CAS  Google Scholar 

  53. Gopal PK, Shepherd MG, Sullivan PA (1984) Analysis of wall glucans from yeast, hyphal and germ-tube forming cells of Candida albicans. J Gen Microbiol 130:3295–3301

    PubMed  CAS  Google Scholar 

  54. Gorka-Niec W, Perlinska-Lenart U, Zembek P, Palamarczyk G, Kruszewska JS (2010) Influence of sorbitol on protein production and glycosylation and cell wall formation in Trichoderma reesei. Fungal Biol 114:855–862

    Article  PubMed  CAS  Google Scholar 

  55. Bartnicki-Garcia S (1999) Glucans, walls, and morphogenesis: On the contributions of J. G. H. Wessels to the golden decades of fungal physiology and beyond. Fungal Genet Biol 27:119–127

    Article  PubMed  CAS  Google Scholar 

  56. Perez P, Garcia-Acha I, Duran A (1983) Effect of papulacandin B on the cell wall and growth of Geotrichum lactis. J Gen Microbiol 129:245–250

    PubMed  CAS  Google Scholar 

  57. Sietsma JH, Wessels JG (1977) Chemical analysis of the hyphal wall of Schizophyllum commune. Biochim Biophys Acta 496:225–239

    Article  PubMed  CAS  Google Scholar 

  58. Tomazett PK, Felix CR, Lenzi HL, de Paula Faria F, de Almeida Soares CM, Pereira M (2010) 1,3-b-D-Glucan synthase of Paracoccidioides brasiliensis: recombinant protein, expression and cytolocalization in the yeast and mycelium phases. Fungal Biol 114:809–816

    Article  PubMed  CAS  Google Scholar 

  59. Latge JP (2010) Tasting the fungal cell wall. Cell Microbiol 12:863–872

    Article  PubMed  CAS  Google Scholar 

  60. Manners DJ, Meyer MT (1977) The molecular structures of some glucans from the cell walls of Schizosaccharomyces pombe. Carbohydr Res 57:189–203

    Article  CAS  Google Scholar 

  61. Magnelli PE, Cipollo JF, Robbins PW (2005) A glucanase-driven fractionation allows redefinition of Schizosaccharomyces pombe cell wall composition and structure: assignment of diglucan. Anal Biochem 336:202–212

    Article  PubMed  CAS  Google Scholar 

  62. Sugawara T, Takahashi S, Osumi M, Ohno N (2004) Refinement of the structures of cell-wall glucans of Schizosaccharomyces pombe by chemical modification and NMR spectroscopy. Carbohydr Res 339: 2255–2265

    Article  PubMed  CAS  Google Scholar 

  63. Magnelli P, Cipollo JF, Abeijon C (2002) A refined method for the determination of Saccharomyces cerevisiae cell wall composition and b-1,6-glucan fine structure. Anal Biochem 301:136–150

    Article  PubMed  CAS  Google Scholar 

  64. Sugawara T, Sato M, Takagi T, Kamasaki T, Ohno N, Osumi M (2003) In situ localization of cell wall a-1,3-glucan in the fission yeast Schizosaccharomyces pombe. J Electron Microsc (Tokyo) 52:237–242

    Article  CAS  Google Scholar 

  65. Bush DA, Horisberger M, Horman I, Wursch P (1974) The wall structure of Schizosaccharomyces pombe. J Gen Microbiol 81:199–206

    Article  PubMed  CAS  Google Scholar 

  66. Cabib E, Durán A (2005) Synthase III-dependent chitin is bound to different acceptors depending on location on the cell wall of budding yeast. J Biol Chem 280:9170–9179

    Article  PubMed  CAS  Google Scholar 

  67. Manners DJ, Masson AJ, Patterson JC (1973) The structure of a b-(1-3)-D-glucan from yeast cell walls. Biochem J 135:19–30

    PubMed  CAS  Google Scholar 

  68. Manners DJ, Masson AJ, Patterson JC, Bjorndal H, Lindberg B (1973) The structure of a b-(1-6)-D-glucan from yeast cell walls. Biochem J 135:31–36

    PubMed  CAS  Google Scholar 

  69. Fontaine T, Simenel C, Dubreucq G, Adam O, Delepierre M, Lemoine J et al (2000) Molecular organization of the alkali-insoluble fraction of Aspergillus fumigatus cell wall. J Biol Chem 275:27594–27607

    PubMed  CAS  Google Scholar 

  70. Montijn RC, van Rinsum J, van Schagen FA, Klis FM (1994) Glucomannoproteins in the cell wall of Saccharomyces cerevisiae contain a novel type of carbohydrate side chain. J Biol Chem 269:19338–19342

    PubMed  CAS  Google Scholar 

  71. Bulawa CE, Slater M, Cabib E, Au-Young J, Sburlati A, Adair WL Jr, Robbins PW (1986) The S. cerevisiae structural gene for chitin synthase is not required for chitin synthesis in vivo. Cell 46:213–225

    Article  PubMed  CAS  Google Scholar 

  72. Reissig JL, Storminger JL, Leloir LF (1955) A modified colorimetric method for the estimation of N-acetylamino sugars. J Biol Chem 217:959–966

    PubMed  CAS  Google Scholar 

  73. Boone C, Sommer SS, Hensel A, Bussey H (1990) Yeast KRE genes provide evidence for a pathway of cell wall b-glucan assembly. J Cell Biol 110:1833–1843

    Article  PubMed  CAS  Google Scholar 

  74. Dijkgraaf GJ, Brown JL, Bussey H (1996) The KNH1 gene of Saccharomyces cerevisiae is a functional homolog of KRE9. Yeast 12:683–692

    Article  PubMed  CAS  Google Scholar 

  75. Dijkgraaf GJ, Abe M, Ohya Y, Bussey H (2002) Mutations in Fks1p affect the cell wall content of b-1,3- and b-1,6-glucan in Saccharomyces cerevisiae. Yeast 19:671–690

    Article  PubMed  CAS  Google Scholar 

  76. Badin J, Jackson C, Schubert M (1953) Improved method for determination of plasma polysaccharides with tryptophan. Proc Soc Exp Biol Med 84:289–291

    PubMed  CAS  Google Scholar 

  77. McKelvy JF, Lee YC (1969) Microheterogeneity of the carbohydrate group of Aspergillus oryzae a-amylase. Arch Biochem Biophys 132:99–110

    Article  PubMed  CAS  Google Scholar 

  78. Lussier M, Sdicu AM, Shahinian S, Bussey H (1998) The Candida albicans KRE9 gene is required for cell wall b-1,6-glucan synthesis and is essential for growth on glucose. Proc Natl Acad Sci U S A 95:9825–9830

    Article  PubMed  CAS  Google Scholar 

  79. Ecker M, Deutzmann R, Lehle L, Mrsa V, Tanner W (2006) Pir proteins of Saccharomyces cerevisiae are attached to b-1,3-glucan by a new protein-carbohydrate linkage. J Biol Chem 281:11523–11529

    Article  PubMed  CAS  Google Scholar 

  80. de Groot PW, Ram AF, Klis FM (2005) Features and functions of covalently linked proteins in fungal cell walls. Fungal Genet Biol 42:657–675

    Article  PubMed  CAS  Google Scholar 

  81. de Groot PW, Yin QY, Weig M, Sosinska GJ, Klis FM, de Koster CG (2007) Mass spectrometric identification of covalently bound cell wall proteins from the fission yeast Schizosaccharomyces pombe. Yeast 24:267–278

    Article  PubMed  CAS  Google Scholar 

  82. Klis FM, Brul S, De Groot PW (2010) Covalently linked wall proteins in ascomycetous fungi. Yeast 27:489–493

    Article  PubMed  CAS  Google Scholar 

  83. Mrsa V, Ecker M, Strahl-Bolsinger S, Nimtz M, Lehle L, Tanner W (1999) Deletion of new covalently linked cell wall glycoproteins alters the electrophoretic mobility of phosphorylated wall components of Saccharomyces cerevisiae. J Bacteriol 181:3076–3086

    PubMed  CAS  Google Scholar 

  84. de Groot PW, de Boer AD, Cunningham J, Dekker HL, de Jong L, Hellingwerf KJ et al (2004) Proteomic analysis of Candida albicans cell walls reveals covalently bound carbohydrate-active enzymes and adhesins. Eukaryot Cell 3:955–965

    Article  PubMed  CAS  Google Scholar 

  85. Carotti C, Ragni E, Palomares O, Fontaine T, Tedeschi G, Rodriguez R et al (2004) Characterization of recombinant forms of the yeast Gas1 protein and identification of residues essential for glucanosyltransferase activity and folding. Eur J Biochem 271:3635–3645

    Article  PubMed  CAS  Google Scholar 

  86. Kapteyn JC, ter Riet B, Vink E, Blad S, De Nobel H, Van Den Ende H et al (2001) Low external pH induces HOG1-dependent changes in the organization of the Saccharomyces cerevisiae cell wall. Mol Microbiol 39:469–479

    Article  PubMed  CAS  Google Scholar 

  87. Mrsa V, Seidl T, Gentzsch M, Tanner W (1997) Specific labelling of cell wall proteins by biotinylation. Identification of four covalently linked O-mannosylated proteins of Saccharomyces cerevisiae. Yeast 13:1145–1154

    Article  PubMed  CAS  Google Scholar 

  88. Mrsa V, Tanner W (1999) Role of NaOH-extractable cell wall proteins Ccw5p, Ccw6p, Ccw7p and Ccw8p (members of the Pir protein family) in stability of the Saccharomyces cerevisiae cell wall. Yeast 15:813–820

    Article  PubMed  CAS  Google Scholar 

  89. Ballou L, Hernandez LM, Alvarado E, Ballou CE (1990) Revision of the oligosaccharide structures of yeast carboxypeptidase Y. Proc Natl Acad Sci U S A 87:3368–3372

    Article  PubMed  CAS  Google Scholar 

  90. Calonge TM, Nakano K, Arellano M, Arai R, Katayama S, Toda T et al (2000) Schizosaccharomyces pombe rho2p GTPase regulates cell wall a-glucan biosynthesis through the protein kinase pck2p. Mol Biol Cell 11:4393–4401

    PubMed  CAS  Google Scholar 

  91. Fuglsang CC, Berka RM, Wahleithner JA, Kauppinen S, Shuster JR, Rasmussen G et al (2000) Biochemical analysis of recombinant fungal mutanases. A new family of a1,3-glucanases with novel carbohydrate-binding domains. J Biol Chem 275:2009–2018

    Article  PubMed  CAS  Google Scholar 

  92. Cortés JC, Konomi M, Martins IM, Munoz J, Moreno MB, Osumi M et al (2007) The (1,3)b-D-glucan synthase subunit Bgs1p is responsible for the fission yeast primary septum formation. Mol Microbiol 65:201–217

    Article  PubMed  CAS  Google Scholar 

  93. Bal AM (2010) The echinocandins: three useful choices or three too many? Int J Antimicrob Agents 35:13–18

    Article  PubMed  CAS  Google Scholar 

  94. Chapman SW, Sullivan DC, Cleary JD (2008) In search of the holy grail of antifungal therapy. Trans Am Clin Climatol Assoc 119:197–215; discussion 215–216

    Google Scholar 

  95. Shao PL, Huang LM, Hsueh PR (2007) Recent advances and challenges in the treatment of invasive fungal infections. Int J Antimicrob Agents 30:487–495

    Article  PubMed  CAS  Google Scholar 

  96. Vicente MF, Basilio A, Cabello A, Pelaez F (2003) Microbial natural products as a source of antifungals. Clin Microbiol Infect 9:15–32

    Article  PubMed  CAS  Google Scholar 

  97. Varona R, Pérez P, Durán A (1983) Effect of papulacandin B on b-glucan synthesis in Schizosaccharomyces pombe. FEMS Microbiol Lett 20:243–247

    CAS  Google Scholar 

  98. Kitamura A, Higuchi S, Hata M, Kawakami K, Yoshida K, Namba K et al (2009) Effect of b-1,6-glucan inhibitors on the invasion process of Candida albicans: potential mechanism of their in vivo efficacy. Antimicrob Agents Chemother 53:3963–3971

    Article  PubMed  CAS  Google Scholar 

  99. Kitamura A, Someya K, Hata M, Nakajima R, Takemura M (2009) Discovery of a small-molecule inhibitor of b-1,6-glucan synthesis. Antimicrob Agents Chemother 53:670–677

    Article  PubMed  CAS  Google Scholar 

  100. Biely P, Kovarik J, Bauer S (1973) Cell wall formation in yeast. An electron microscopic autoradiographic study. Arch Microbiol 94:356–371

    CAS  Google Scholar 

  101. Feldmesser M, Kress Y, Mednick A, Casadevall A (2000) The effect of the echinocandin analogue caspofungin on cell wall glucan synthesis by Cryptococcus neoformans. J Infect Dis 182:1791–1795

    Article  PubMed  CAS  Google Scholar 

  102. Johnson BF, Yoo BY, Calleja GB (1973) Cell division in yeasts: movement of organelles associated with cell plate growth of Schizosaccharomyces pombe. J Bacteriol 115:358–366

    PubMed  CAS  Google Scholar 

  103. Osumi M, Sato M, Ishijima SA, Konomi M, Takagi T, Yaguchi H (1998) Dynamics of cell wall formation in fission yeast, Schizosaccharomyces pombe. Fungal Genet Biol 24:178–206

    Article  PubMed  CAS  Google Scholar 

  104. Roh DH, Bowers B, Schmidt M, Cabib E (2002) The septation apparatus, an autonomous system in budding yeast. Mol Biol Cell 13:2747–2759

    Article  PubMed  CAS  Google Scholar 

  105. Schmidt M, Bowers B, Varma A, Roh DH, Cabib E (2002) In budding yeast, contraction of the actomyosin ring and formation of the primary septum at cytokinesis depend on each other. J Cell Sci 115:293–302

    PubMed  CAS  Google Scholar 

  106. Coluccio A, Bogengruber E, Conrad MN, Dresser ME, Briza P, Neiman AM (2004) Morphogenetic pathway of spore wall assembly in Saccharomyces cerevisiae. Eukaryot Cell 3:1464–1475

    Article  PubMed  CAS  Google Scholar 

  107. Bowers B, Levin G, Cabib E (1974) Effect of polyoxin D on chitin synthesis and septum formation in Saccharomyces cerevisiae. J Bacteriol 119:564–575

    PubMed  CAS  Google Scholar 

  108. Cabib E, Sburlati A, Bowers B, Silverman SJ (1989) Chitin synthase 1, an auxiliary enzyme for chitin synthesis in Saccharomyces cerevisiae. J Cell Biol 108:1665–1672

    Article  PubMed  CAS  Google Scholar 

  109. Shaw JA, Mol PC, Bowers B, Silverman SJ, Valdivieso MH, Durán A et al (1991) The function of chitin synthases 2 and 3 in the Saccharomyces cerevisiae cell cycle. J Cell Biol 114:111–123

    Article  PubMed  CAS  Google Scholar 

  110. Konomi M, Fujimoto K, Toda T, Osumi M (2003) Characterization and behaviour of a-glucan synthase in Schizosaccharomyces pombe as revealed by electron microscopy. Yeast 20:427–438

    Article  PubMed  CAS  Google Scholar 

  111. Osumi M, Yamada N, Kobori H, Taki A, Naito N, Baba M et al (1989) Cell wall formation in regenerating protoplasts of Schizosaccharomyces pombe: study by high resolution, low voltage scanning electron microscopy. J Electron Microsc (Tokyo) 38:457–468

    CAS  Google Scholar 

  112. Konomi M, Ishiguro J, Osumi M (2000) Abnormal formation of the glucan network from regenerating protoplasts in Schizosaccharomyces pombe cps8 actin point mutant. J Electron Microsc (Tokyo) 49:569–578

    Article  CAS  Google Scholar 

  113. Osumi M (1998) The ultrastructure of yeast: cell wall structure and formation. Micron 29:207–233

    Article  PubMed  CAS  Google Scholar 

  114. Kopecka M, Fleet GH, Phaff HJ (1995) Ultrastructure of the cell wall of Schizosaccharomyces pombe following treatment with various glucanases. J Struct Biol 114:140–152

    Article  PubMed  CAS  Google Scholar 

  115. Sipiczki M, Yamaguchi M, Grallert A, Takeo K, Zilahi E, Bozsik A et al (2000) Role of cell shape in determination of the division plane in Schizosaccharomyces pombe: random orientation of septa in spherical cells. J Bacteriol 182:1693–1701

    Article  PubMed  CAS  Google Scholar 

  116. Osumi M, Yamada N, Yaguchi H, Kobori H, Nagatani T, Sato M (1995) Ultrahigh-resolution low-voltage SEM reveals ultrastructure of the glucan network formation from fission yeast protoplast. J Electron Microsc (Tokyo) 44:198–206

    CAS  Google Scholar 

  117. Kobori H, Yamada N, Taki A, Osumi M (1989) Actin is associated with the formation of the cell wall in reverting protoplasts of the fission yeast Schizosaccharomyces pombe. J Cell Sci 94:635–646

    PubMed  Google Scholar 

  118. Bernard M, Latge JP (2001) Aspergillus fumigatus cell wall: composition and biosynthesis. Med Mycol 39:9–17

    PubMed  CAS  Google Scholar 

  119. Dufrene YF (2010) Atomic force microscopy of fungal cell walls: an update. Yeast 27:465–471

    Article  PubMed  CAS  Google Scholar 

  120. Zhao L, Schaefer D, Xu H, Modi SJ, LaCourse WR, Marten MR (2005) Elastic properties of the cell wall of Aspergillus nidulans studied with atomic force microscopy. Biotechnol Prog 21:292–299

    Article  PubMed  CAS  Google Scholar 

  121. Osumi M, Konomi M, Sugawara T, Takagi T, Baba M (2006) High-pressure freezing is a powerful tool for visualization of Schizosaccharomyces pombe cells: ultra-low temperature and low-voltage scanning electron microscopy and immunoelectron microscopy. J Electron Microsc (Tokyo) 55:75–88

    Article  CAS  Google Scholar 

  122. Humbel BM, Konomi M, Takagi T, Kamasawa N, Ishijima SA, Osumi M (2001) In situ localization of b-glucans in the cell wall of Schizosaccharomyces pombe. Yeast 18:433–444

    Article  PubMed  CAS  Google Scholar 

  123. Meikle PJ, Bonig I, Hoogenraad NJ, Clarke AE, Stone BA (1991) The location of (1-3)-b-glucans in the walls of pollen tubes of Nicotiana alata using a (1-3)-b-glucan-specific monoclonal antibody. Planta 185:1–8

    Article  CAS  Google Scholar 

  124. Meikle PJ, Hoogenraad NJ, Bonig I, Clarke AE, Stone BA (1994) A (1->3,1->4)-beta-glucan-specific monoclonal antibody and its use in the quantitation and immunocytochemical location of (1->3,1->4)-beta-glucans. Plant J 5:1–9

    Article  PubMed  CAS  Google Scholar 

  125. Montijn RC, Vink E, Muller WH, Verkleij AJ, Van Den Ende H, Henrissat B et al (1999) Localization of synthesis of b1,6-glucan in Saccharomyces cerevisiae. J Bacteriol 181:7414–7420

    PubMed  CAS  Google Scholar 

  126. Adachi Y, Ohno N, Yadomae T (1994) Preparation and antigen specificity of an anti-(1-3)-b-D-glucan antibody. Biol Pharm Bull 17:1508–1512

    Article  PubMed  CAS  Google Scholar 

  127. Reese AJ, Yoneda A, Breger JA, Beauvais A, Liu H, Griffith CL et al (2007) Loss of cell wall a(1-3) ­glucan affects Cryptococcus neoformans from ultrastructure to virulence. Mol Microbiol 63:1385–1398

    Article  PubMed  CAS  Google Scholar 

  128. Rappleye CA, Eissenberg LG, Goldman WE (2007) Histoplasma capsulatum a-(1,3)-glucan blocks innate immune recognition by the b-glucan receptor. Proc Natl Acad Sci U S A 104:1366–1370

    Article  PubMed  CAS  Google Scholar 

  129. Eissenberg LG, Moser SA, Goldman WE (1997) Alterations to the cell wall of Histoplasma capsulatum yeasts during infection of macrophages or epithelial cells. J Infect Dis 175:1538–1544

    Article  PubMed  CAS  Google Scholar 

  130. Kugler S, Schurtz Sebghati T, Groppe Eissenberg L, Goldman WE (2000) Phenotypic variation and intracellular parasitism by Histoplasma capsulatum. Proc Natl Acad Sci U S A 97:8794–8798

    Article  PubMed  CAS  Google Scholar 

  131. Gonzalez M, Goddard N, Hicks C, Ovalle R, Rauceo JM, Jue CK et al (2010) A screen for deficiencies in GPI-anchorage of wall glycoproteins in yeast. Yeast 27:583–596

    Article  PubMed  CAS  Google Scholar 

  132. Cabib E, Bowers B, Roberts RL (1983) Vectorial synthesis of a polysaccharide by isolated plasma membranes. Proc Natl Acad Sci U S A 80:3318–3321

    Article  PubMed  CAS  Google Scholar 

  133. Molano J, Bowers B, Cabib E (1980) Distribution of chitin in the yeast cell wall. An ultrastructural and chemical study. J Cell Biol 85:199–212

    Article  PubMed  CAS  Google Scholar 

  134. Roberts RL, Bowers B, Slater ML, Cabib E (1983) Chitin synthesis and localization in cell division cycle mutants of Saccharomyces cerevisiae. Mol Cell Biol 3:922–930

    PubMed  CAS  Google Scholar 

Download references

Acknowledgements

We thank D. Posner for language revision. This work was supported by grants BFU2010-15641 and BIO2009-10597 from the Dirección General de Investigación, MICINN, Spain, and grant CSI038A11-2 from the Junta de Castilla y León, Spain.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Pilar Pérez .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2013 Springer Science+Business Media, LLC

About this chapter

Cite this chapter

Pérez, P., Ribas, J.C. (2013). Fungal Cell Wall Analysis. In: Gupta, V., Tuohy, M., Ayyachamy, M., Turner, K., O’Donovan, A. (eds) Laboratory Protocols in Fungal Biology. Fungal Biology. Springer, New York, NY. https://doi.org/10.1007/978-1-4614-2356-0_12

Download citation

Publish with us

Policies and ethics