Abstract
Methods and procedures in molecular biology used to study fungal pathogenesis have significantly improved during the last decade. In this chapter, we provide step-by-step procedures for performing genetics and biochemical studies in the human pathogenic fungal microorganism Cryptococcus neoformans (Cn). These methods are employed for studying the pathobiology of Cn and for experimental validation of theoretical models of fungal pathogenicity.
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References
Harrison, T. S. (2009) The burden of HIV-associated cryptococcal disease. AIDS 23, 531–2.
Park, B. J., Wannemuehler, K. A., Marston, B. J., Govender, N., Pappas, P. G., and Chiller, T. M. (2009) Estimation of the current global burden of cryptococcal meningitis among persons living with HIV/AIDS. AIDS 23, 525–30.
Hajjeh, R. A., Conn, L. A., Stephens, D. S., Baughman, W., Hamill, R., Graviss, E., Pappas, P. G., Thomas, C., Reingold, A., Rothrock, G., Hutwagner, L. C., Schuchat, A., Brandt, M. E., and Pinner, R. W. (1999) Cryptococcosis: population-based multistate active surveillance and risk factors in human immunodeficiency virus-infected persons. Cryptococcal Active Surveillance Group. J Infect Dis 179, 449–54.
Kaplan, M. H., Rosen, P. P., and Armstrong, D. (1977) Cryptococcosis in a cancer hospital: clinical and pathological correlates in forty-six patients. Cancer 39, 2265–74.
White, M., Cirrincione, C., Blevins, A., and Armstrong, D. (1992) Cryptococcal meningitis: outcome in patients with AIDS and patients with neoplastic disease. J Infect Dis 165, 960–3.
Kohno, S., Varma, A., Kwon-Chung, K. J., and Hara, K. (1994) Epidemiology studies of clinical isolates of Cryptococcus neoformans of Japan by restriction fragment length polymorphism. Kansenshogaku Zasshi 68, 1512–7.
Shaariah, W., Morad, Z., and Suleiman, A. B. (1992) Cryptococcosis in renal transplant recipients. Transplant Proc 24, 1898–9.
Husain, S., Wagener, M. M., and Singh, N. (2001) Cryptococcus neoformans infection in organ transplant recipients: variables influencing clinical characteristics and outcome. Emerg Infect Dis 7, 375–81.
Fraser, J. A., Giles, S. S., Wenink, E. C., Geunes-Boyer, S. G., Wright, J. R., Diezmann, S., Allen, A., Stajich, J. E., Dietrich, F. S., Perfect, J. R., and Heitman, J. (2005) Same-sex mating and the origin of the Vancouver Island Cryptococcus gattii outbreak. Nature 437, 1360–4.
Seaton, R. A., Verma, N., Naraqi, S., Wembri, J. P., and Warrell, D. A. (1997) Visual loss in immunocompetent patients with Cryptococcus neoformans var. gattii meningitis. Trans R Soc Trop Med Hyg 91, 44–9.
Seaton, R. A., Naraqi, S., Wembri, J. P., and Warrell, D. A. (1996) Predictors of outcome in Cryptococcus neoformans var. gattii meningitis. Qjm 89, 423–8.
Findley, K., Rodriguez-Carres, M., Metin, B., Kroiss, J., Fonseca, A., Vilgalys, R., and Heitman, J. (2009) Phylogeny and phenotypic characterization of pathogenic Cryptococcus species and closely related saprobic taxa in the Tremellales. Eukaryot Cell 8, 353–61.
Perfect, J. R. (2005) Cryptococcus neoformans: a sugar-coated killer with designer genes. FEMS Immunol Med Microbiol 45, 395–404.
Casadevall, A., and Perfect, J. R. (1998) Cryptococcus neoformans, ASM Press, Washington, DC, 381–405.
Hull, C. M., and Heitman, J. (2002) Genetics of Cryptococcus neoformans. Annu Rev Genet 36, 557–615.
Toffaletti, D. L., Rude, T. H., Johnston, S. A., Durack, D. T., and Perfect, J. R. (1993) Gene transfer in Cryptococcus neoformans by use of biolistic delivery of DNA. J. Bacteriol. 175, 1405–11.
Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular cloning: a laboratory manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.
Baker, L. G., Specht, C. A., Donlin, M. J., and Lodge, J. K. (2007) Chitosan, the deacetylated form of chitin, is necessary for cell wall integrity in Cryptococcus neoformans. Eukaryot Cell 6, 855–67.
Mandala, S. M., Thornton, R. A., Frommer, B. R., and et al. (1995) The discovery of australifungin, a novel inhibitor of sphinganine N-acyltransferase from Sporormiella australis. Producing organism, fermentation, isolation, and biological activity. J. Antibiot. (Tokyo) 48, 349–56.
Luberto, C., Toffaletti, D. L., Wills, E. A., Tucker, S. C., Casadevall, A., Perfect, J. R., Hannun, Y. A., and Del Poeta, M. (2001) Roles for inositol-phosphoryl ceramide synthase 1 (IPC1) in pathogenesis of C. neoformans. Genes Dev. 15, 201–12.
Fischl, A. S., Liu, Y., Browdy, A., and Cremesti, A. E. (2000) Inositolphosphoryl ceramide synthase from yeast. Methods Enzymol. 311, 123–30.
Bielawski, J., Pierce, J. S., Snider, J., Rembiesa, B., Szulc, Z. M., and Bielawska, A. (2009) Comprehensive quantitative analysis of bioactive sphingolipids by high-performance liquid chromatography-tandem mass spectrometry. Methods Mol Biol 579, 443–67.
Bielawski, J., Szulc, Z. M., Hannun, Y. A., and Bielawska, A. (2006) Simultaneous quantitative analysis of bioactive sphingolipids by high-performance liquid chromatography-tandem mass spectrometry. Methods 39, 82–91.
Hanson, B. A., and Lester, R. L. (1980) The extraction of inositol-containing phospholipids and phosphatidylcholine from Saccharomyces cerevisiae and Neurospora crassa. J Lipid Res 21, 309–15.
Acknowledgments
This work was supported by Grants AI56168 and AI72142 (to M.D.P) and was conducted in a facility constructed with support from the National Institutes of Health, Grant Number C06 RR015455 from the Extramural Research Facilities Program of the National Center for Research Resources. Dr. Maurizio Del Poeta is a Burroughs Wellcome New Investigator in Pathogenesis of Infectious Diseases.
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Singh, A., Qureshi, A., Del Poeta, M. (2011). Quantitation of Cellular Components in Cryptococcus neoformans for System Biology Analysis. In: Becskei, A. (eds) Yeast Genetic Networks. Methods in Molecular Biology, vol 734. Humana Press. https://doi.org/10.1007/978-1-61779-086-7_16
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DOI: https://doi.org/10.1007/978-1-61779-086-7_16
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