Fungal kinases and transcription factors regulating brain infection in Cryptococcus neoformans.
Animals
Blood-Brain Barrier
/ metabolism
Brain
/ microbiology
Cryptococcus gattii
/ genetics
Cryptococcus neoformans
/ genetics
Disease Models, Animal
Female
Fungal Proteins
Gene Expression Profiling
Gene Expression Regulation, Fungal
Homeodomain Proteins
/ genetics
Humans
Meningitis, Cryptococcal
/ microbiology
Meningoencephalitis
/ microbiology
Mice
Mutagenesis
Mutation
Permeability
Phosphotransferases
/ genetics
Signal Transduction
/ genetics
Transcription Factors
/ genetics
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
23 03 2020
23 03 2020
Historique:
received:
19
06
2019
accepted:
03
03
2020
entrez:
7
4
2020
pubmed:
7
4
2020
medline:
22
7
2020
Statut:
epublish
Résumé
Cryptococcus neoformans causes fatal fungal meningoencephalitis. Here, we study the roles played by fungal kinases and transcription factors (TFs) in blood-brain barrier (BBB) crossing and brain infection in mice. We use a brain infectivity assay to screen signature-tagged mutagenesis (STM)-based libraries of mutants defective in kinases and TFs, generated in the C. neoformans H99 strain. We also monitor in vivo transcription profiles of kinases and TFs during host infection using NanoString technology. These analyses identify signalling components involved in BBB adhesion and crossing, or survival in the brain parenchyma. The TFs Pdr802, Hob1, and Sre1 are required for infection under all the conditions tested here. Hob1 controls the expression of several factors involved in brain infection, including inositol transporters, a metalloprotease, PDR802, and SRE1. However, Hob1 is dispensable for most cellular functions in Cryptococcus deuterogattii R265, a strain that does not target the brain during infection. Our results indicate that Hob1 is a master regulator of brain infectivity in C. neoformans.
Identifiants
pubmed: 32251295
doi: 10.1038/s41467-020-15329-2
pii: 10.1038/s41467-020-15329-2
pmc: PMC7090016
doi:
Substances chimiques
Fungal Proteins
0
Homeodomain Proteins
0
Transcription Factors
0
Phosphotransferases
EC 2.7.-
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1521Références
Fisher, M. C. et al. Emerging fungal threats to animal, plant and ecosystem health. Nature 484, 186–194 (2012).
pubmed: 22498624
doi: 10.1038/nature10947
pmcid: 22498624
Brown, G. D. et al. Hidden killers: human fungal infections. Sci. Transl. Med. 4, 165rv113 (2012).
doi: 10.1126/scitranslmed.3004404
Rajasingham, R. et al. Global burden of disease of HIV-associated cryptococcal meningitis: an updated analysis. Lancet Infect. Dis. 17, 873–881 (2017).
pubmed: 28483415
pmcid: 5818156
doi: 10.1016/S1473-3099(17)30243-8
Kwon-Chung, K. J. et al. The case for adopting the “Species Complex” nomenclature for the etiologic agents of cryptococcosis. mSphere 2, e00357–16 (2017).
pubmed: 28101535
pmcid: 5227069
doi: 10.1128/mSphere.00357-16
Zaragoza, O. et al. The capsule of the fungal pathogen Cryptococcus neoformans. Adv. Appl. Microbiol. 68, 133–216 (2009).
pubmed: 19426855
pmcid: 2739887
doi: 10.1016/S0065-2164(09)01204-0
Bojarczuk, A. et al. Cryptococcus neoformans intracellular proliferation and capsule size determines early macrophage control of infection. Sci. Rep. 6, 21489 (2016).
pubmed: 26887656
pmcid: 4757829
doi: 10.1038/srep21489
Wang, Y., Aisen, P. & Casadevall, A. Cryptococcus neoformans melanin and virulence: mechanism of action. Infect. Immun. 63, 3131–3136 (1995).
pubmed: 7622240
pmcid: 173427
doi: 10.1128/IAI.63.8.3131-3136.1995
Mednick, A. J., Nosanchuk, J. D. & Casadevall, A. Melanization of Cryptococcus neoformans affects lung inflammatory responses during cryptococcal infection. Infect. Immun. 73, 2012–2019 (2005).
pubmed: 15784542
pmcid: 1087470
doi: 10.1128/IAI.73.4.2012-2019.2005
Ma, H., Croudace, J. E., Lammas, D. A. & May, R. C. Expulsion of live pathogenic yeast by macrophages. Curr. Biol. 16, 2156–2160 (2006).
pubmed: 17084701
doi: 10.1016/j.cub.2006.09.032
pmcid: 17084701
Johnston, S. A. & May, R. C. Cryptococcus interactions with macrophages: evasion and manipulation of the phagosome by a fungal pathogen. Cell Microbiol. 15, 403–411 (2013).
pubmed: 23127124
doi: 10.1111/cmi.12067
pmcid: 23127124
Johnston, S. A. & May, R. C. The human fungal pathogen Cryptococcus neoformans escapes macrophages by a phagosome emptying mechanism that is inhibited by Arp2/3 complex-mediated actin polymerisation. PLoS Pathog. 6, e1001041 (2010).
pubmed: 20714349
pmcid: 2920849
doi: 10.1371/journal.ppat.1001041
Dziegielewska, K. M., Ek, J., Habgood, M. D. & Saunders, N. R. Development of the choroid plexus. Microsc. Res. Tech. 52, 5–20 (2001).
pubmed: 11135444
doi: 10.1002/1097-0029(20010101)52:1<5::AID-JEMT3>3.0.CO;2-J
pmcid: 11135444
Esher, S. K., Zaragoza, O. & Alspaugh, J. A. Cryptococcal pathogenic mechanisms: a dangerous trip from the environment to the brain. Mem. Inst. Oswaldo Cruz 113, e180057 (2018).
pubmed: 29668825
pmcid: 5909089
doi: 10.1590/0074-02760180057
Griffiths, E. J., Kretschmer, M. & Kronstad, J. W. Aimless mutants of Cryptococcus neoformans: failure to disseminate. Fungal Biol. Rev. 26, 61–72 (2012).
pubmed: 23189087
pmcid: 3505455
doi: 10.1016/j.fbr.2012.02.004
Lee, K. T. et al. Systematic functional analysis of kinases in the fungal pathogen Cryptococcus neoformans. Nat. Commun. 7, 12766 (2016).
pubmed: 27677328
pmcid: 5052723
doi: 10.1038/ncomms12766
Jung, K. W. et al. Systematic functional profiling of transcription factor networks in Cryptococcus neoformans. Nat. Commun. 6, 6757 (2015).
pubmed: 25849373
pmcid: 4391232
doi: 10.1038/ncomms7757
Bahn, Y. S., Hicks, J. K., Giles, S. S., Cox, G. M. & Heitman, J. Adenylyl cyclase-associated protein Aca1 regulates virulence and differentiation of Cryptococcus neoformans via the cyclic AMP-protein kinase A cascade. Eukaryot. Cell 3, 1476–1491 (2004).
pubmed: 15590822
pmcid: 539029
doi: 10.1128/EC.3.6.1476-1491.2004
Alspaugh, J. A. et al. Adenylyl cyclase functions downstream of the Galpha protein Gpa1 and controls mating and pathogenicity of Cryptococcus neoformans. Eukaryot. Cell 1, 75–84 (2002).
pubmed: 12455973
pmcid: 118042
doi: 10.1128/EC.1.1.75-84.2002
Bahn, Y. S., Geunes-Boyer, S. & Heitman, J. Ssk2 mitogen-activated protein kinase kinase kinase governs divergent patterns of the stress-activated Hog1 signaling pathway in Cryptococcus neoformans. Eukaryot. Cell 6, 2278–2289 (2007).
pubmed: 17951522
pmcid: 2168243
doi: 10.1128/EC.00349-07
Bahn, Y. S., Kojima, K., Cox, G. M. & Heitman, J. Specialization of the HOG pathway and its impact on differentiation and virulence of Cryptococcus neoformans. Mol. Biol. Cell 16, 2285–2300 (2005).
pubmed: 15728721
pmcid: 1087235
doi: 10.1091/mbc.e04-11-0987
Gerik, K. J. et al. Cell wall integrity is dependent on the PKC1 signal transduction pathway in Cryptococcus neoformans. Mol. Microbiol. 58, 393–408 (2005).
pubmed: 16194228
doi: 10.1111/j.1365-2958.2005.04843.x
pmcid: 16194228
Kraus, P. R., Fox, D. S., Cox, G. M. & Heitman, J. The Cryptococcus neoformans MAP kinase Mpk1 regulates cell integrity in response to antifungal drugs and loss of calcineurin function. Mol. Microbiol. 48, 1377–1387 (2003).
pubmed: 12787363
pmcid: 1635492
doi: 10.1046/j.1365-2958.2003.03508.x
Cheon, S. A. et al. Unique evolution of the UPR pathway with a novel bZIP transcription factor, Hxl1, for controlling pathogenicity of Cryptococcus neoformans. PLoS Pathog. 7, e1002177 (2011).
pubmed: 21852949
pmcid: 3154848
doi: 10.1371/journal.ppat.1002177
Liu, O. W. et al. Systematic genetic analysis of virulence in the human fungal pathogen Cryptococcus neoformans. Cell 135, 174–188 (2008).
pubmed: 18854164
pmcid: 2628477
doi: 10.1016/j.cell.2008.07.046
Cramer, K. L., Gerrald, Q. D., Nichols, C. B., Price, M. S. & Alspaugh, J. A. Transcription factor Nrg1 mediates capsule formation, stress response, and pathogenesis in Cryptococcus neoformans. Eukaryot. Cell 5, 1147–1156 (2006).
pubmed: 16835458
pmcid: 1489281
doi: 10.1128/EC.00145-06
Xu, W., Solis, N. V., Filler, S. G. & Mitchell, A. P. Pathogen gene expression profiling during infection using a NanoString nCounter platform. Methods Mol. Biol. 1361, 57–65 (2016).
pubmed: 26483015
pmcid: 5317040
doi: 10.1007/978-1-4939-3079-1_3
Cadieux, B. et al. The mannoprotein Cig1 supports iron acquisition from heme and virulence in the pathogenic fungus Cryptococcus neoformans. J. Infect. Dis. 207, 1339–1347 (2013).
pubmed: 23322859
pmcid: 3603535
doi: 10.1093/infdis/jit029
Jung, W. H., Hu, G., Kuo, W. & Kronstad, J. W. Role of ferroxidases in iron uptake and virulence of Cryptococcus neoformans. Eukaryot. Cell 8, 1511–1520 (2009).
pubmed: 19700638
pmcid: 2756870
doi: 10.1128/EC.00166-09
Lev, S. et al. Pho4 is essential for dissemination of Cryptococcus neoformans to the host brain by promoting phosphate uptake and growth at alkaline pH. mSphere 2, e00381–16 (2017).
pubmed: 28144629
pmcid: 5266496
doi: 10.1128/mSphere.00381-16
Vu, K., Weksler, B., Romero, I., Couraud, P. O. & Gelli, A. Immortalized human brain endothelial cell line hCMEC/D3 as a model of the blood-brain barrier facilitates in vitro studies of central nervous system infection by Cryptococcus neoformans. Eukaryot. Cell 8, 1803–1807 (2009).
pubmed: 19767445
pmcid: 2772405
doi: 10.1128/EC.00240-09
Vu, K. et al. Invasion of the central nervous system by Cryptococcus neoformans requires a secreted fungal metalloprotease. mBio 5, e01101–01114 (2014).
pubmed: 24895304
pmcid: 4049100
doi: 10.1128/mBio.01101-14
Santiago-Tirado, F. H., Onken, M. D., Cooper, J. A., Klein, R. S. & Doering, T. L. Trojan horse transit contributes to blood-brain barrier crossing of a eukaryotic pathogen. mBio 8, e02183–02116 (2017).
pubmed: 28143979
pmcid: 5285505
doi: 10.1128/mBio.02183-16
Ngamskulrungroj, P., Chang, Y., Sionov, E. & Kwon-Chung, K. J. The primary target organ of Cryptococcus gattii is different from that of Cryptococcus neoformans in a murine model. mBio 3, e00103–00112 (2012).
pubmed: 22570277
pmcid: 3350374
doi: 10.1128/mBio.00103-12
Bielska, E. & May, R. C. What makes Cryptococcus gattii a pathogen? FEMS Yeast Res. 16, fov106 (2015).
pubmed: 26614308
doi: 10.1093/femsyr/fov106
pmcid: 26614308
Fraser, J. A. et al. Same-sex mating and the origin of the Vancouver Island Cryptococcus gattii outbreak. Nature 437, 1360–1364 (2005).
pubmed: 16222245
doi: 10.1038/nature04220
pmcid: 16222245
Ngamskulrungroj, P., Price, J., Sorrell, T., Perfect, J. R. & Meyer, W. Cryptococcus gattii virulence composite: candidate genes revealed by microarray analysis of high and less virulent Vancouver island outbreak strains. PLoS ONE 6, e16076 (2011).
pubmed: 21249145
pmcid: 3020960
doi: 10.1371/journal.pone.0016076
Roelants, F. M., Leskoske, K. L., Martinez Marshall, M. N., Locke, M. N. & Thorner, J. The TORC2-dependent signaling network in the yeast Saccharomyces cerevisiae. Biomolecules 7, pii: E66 (2017).
doi: 10.3390/biom7030066
Liu, T. B. et al. Brain inositol is a novel stimulator for promoting Cryptococcus penetration of the blood-brain barrier. PLoS Pathog. 9, e1003247 (2013).
pubmed: 23592982
pmcid: 3617100
doi: 10.1371/journal.ppat.1003247
Jong, A. et al. Identification and characterization of CPS1 as a hyaluronic acid synthase contributing to the pathogenesis of Cryptococcus neoformans infection. Eukaryot. Cell 6, 1486–1496 (2007).
pubmed: 17545316
pmcid: 1951127
doi: 10.1128/EC.00120-07
Cox, G. M., Mukherjee, J., Cole, G. T., Casadevall, A. & Perfect, J. R. Urease as a virulence factor in experimental cryptococcosis. Infect. Immun. 68, 443–448 (2000).
pubmed: 10639402
pmcid: 97161
doi: 10.1128/IAI.68.2.443-448.2000
Olszewski, M. A. et al. Urease expression by Cryptococcus neoformans promotes microvascular sequestration, thereby enhancing central nervous system invasion. Am. J. Pathol. 164, 1761–1771 (2004).
pubmed: 15111322
pmcid: 1615675
doi: 10.1016/S0002-9440(10)63734-0
Cox, G. M. et al. Extracellular phospholipase activity is a virulence factor for Cryptococcus neoformans. Mol. Microbiol. 39, 166–175 (2001).
pubmed: 11123698
doi: 10.1046/j.1365-2958.2001.02236.x
pmcid: 11123698
Tseng, H. K. et al. Identification of genes from the fungal pathogen Cryptococcus neoformans related to transmigration into the central nervous system. PLoS ONE 7, e45083 (2012).
pubmed: 23028773
pmcid: 3447876
doi: 10.1371/journal.pone.0045083
Price, M. S. et al. Cryptococcus neoformans requires a functional glycolytic pathway for disease but not persistence in the host. mBio 2, e00103–00111 (2011).
pubmed: 21652778
pmcid: 3110414
doi: 10.1128/mBio.00103-11
Lee, H., Khanal Lamichhane, A., Garraffo, H. M., Kwon-Chung, K. J. & Chang, Y. C. Involvement of PDK1, PKC and TOR signalling pathways in basal fluconazole tolerance in Cryptococcus neoformans. Mol. Microbiol 84, 130–146 (2012).
pubmed: 22339665
pmcid: 3313003
doi: 10.1111/j.1365-2958.2012.08016.x
Fanning, S. et al. Divergent targets of Candida albicans biofilm regulator Bcr1 in vitro and in vivo. Eukaryot. Cell 11, 896–904 (2012).
pubmed: 22544909
pmcid: 3416506
doi: 10.1128/EC.00103-12
O’Meara, T. R. et al. The Cryptococcus neoformans Rim101 transcription factor directly regulates genes required for adaptation to the host. Mol. Cell Biol. 34, 673–684 (2014).
pubmed: 24324006
pmcid: 3911494
doi: 10.1128/MCB.01359-13
Jung, K. W. et al. Evolutionarily conserved and divergent roles of unfolded protein response (UPR) in the pathogenic Cryptococcus species complex. Sci. Rep. 8, 8132 (2018).
pubmed: 29802329
pmcid: 5970146
doi: 10.1038/s41598-018-26405-5