Amino Acid Permeases and Virulence in Cryptococcus neoformans.
Fungal opportunistic pathogens colonize various environments, from plants and wood to human and animal tissue. Regarding human pathogens, one great challenge during contrasting niche occupation is the adaptation to different conditions, such as temperature, osmolarity, salinity, pressure, oxidative stress and nutritional availability, which may constitute sources of stress that need to be tolerated and overcome. As an opportunistic pathogen, C. neoformans faces exactly these situations during the transition from the environment to the human host, encountering nutritional constraints. Our previous and current research on amino acid biosynthetic pathways indicates that amino acid permeases are regulated by the presence of the amino acids, nitrogen and temperature. Saccharomyces cerevisiae and Candida albicans have twenty-four and twenty-seven genes encoding amino acid permeases, respectively; conversely, they are scarce in number in Basidiomycetes (C. neoformans, Coprinopsis cinerea and Ustilago maydis), where nine to ten permease genes can be found depending on the species. In this study, we have demonstrated that two amino acid permeases are essential for virulence in C. neoformans. Our data showed that C. neoformans uses two global and redundant amino acid permeases, Aap4 and Aap5 to respond correctly to thermal and oxidative stress. Double deletion of these permeases causes growth arrest in C. neoformans at 37°C and in the presence of hydrogen peroxide. The inability to uptake amino acid at a higher temperature and under oxidative stress also led to virulence attenuation in vivo. Our data showed that thermosensitivity caused by the lack of permeases Aap4 and Aap5 can be remedied by alkaline conditions (higher pH) and salinity. Permeases Aap4 and Aap5 are also required during fluconazole stress and they are the target of the plant secondary metabolite eugenol, a potent antifungal inhibitor that targets amino acid permeases. In summary, our work unravels (i) interesting physiological property of C. neoformans regarding its amino acid uptake system; (ii) an important aspect of virulence, which is the need for amino acid permeases during thermal and oxidative stress resistance and, hence, host invasion and colonization; and (iii) provides a convenient prototype for antifungal development, which are the amino acid permeases Aap4/Aap5 and their inhibitor.
Instituto de Ciências Ambientais, Química e Farmacêuticas, Universidade Federal de São Paulo, Rua Arthur Ridel, 275, Diadema, SP, Brazil.
Kevin Felipe Cruz
KF
Martho
Instituto de Ciências Ambientais, Química e Farmacêuticas, Universidade Federal de São Paulo, Rua Arthur Ridel, 275, Diadema, SP, Brazil.
Amanda Teixeira
AT
de Melo
Quantitative Pathology Unit, Adolfo Lutz Institute, São Paulo, Brazil.
Juliana Possato Fernandes
JP
Takahashi
Quantitative Pathology Unit, Adolfo Lutz Institute, São Paulo, Brazil.
Juliana Mariotti
JM
Guerra
Mycology Unit, Adolfo Lutz Institute, Secretary of Health, São Paulo, Brazil.
Dayane Cristina da Silva
DC
Santos
Mycology Unit, Adolfo Lutz Institute, Secretary of Health, São Paulo, Brazil.
Sônia Ueda
SU
Purisco
Mycology Unit, Adolfo Lutz Institute, Secretary of Health, São Paulo, Brazil.
Márcia de Souza Carvalho
MS
Melhem
Mycology Unit, Adolfo Lutz Institute, Secretary of Health, São Paulo, Brazil.
Raquel Dos Anjos
RD
Fazioli
Instituto de Ciências Ambientais, Química e Farmacêuticas, Universidade Federal de São Paulo, Rua Arthur Ridel, 275, Diadema, SP, Brazil.
Clerlune
C
Phanord
Instituto de Ciências Ambientais, Química e Farmacêuticas, Universidade Federal de São Paulo, Rua Arthur Ridel, 275, Diadema, SP, Brazil.
Patrícia
P
Sartorelli
Instituto de Ciências Ambientais, Química e Farmacêuticas, Universidade Federal de São Paulo, Rua Arthur Ridel, 275, Diadema, SP, Brazil.
Marcelo A
MA
Vallim
Instituto de Ciências Ambientais, Química e Farmacêuticas, Universidade Federal de São Paulo, Rua Arthur Ridel, 275, Diadema, SP, Brazil.
Renata C
http://orcid.org/0000-0001-9902-9396
RC
Pascon
D026905:Amino Acid Transport Systems / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D000935:Antifungal Agents / Q000494:pharmacology; D002244:Carbon / Q000378:metabolism; D003453:Cryptococcosis / Q000382:microbiology* / Q000401:mortality; D003455:Cryptococcus neoformans / Q000187:drug effects / Q000201:enzymology* / Q000378:metabolism / Q000472:pathogenicity*; D004195:Disease Models, Animal; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D005947:Glucose / Q000378:metabolism; D008297:Male; D051379:Mice; D008826:Microbial Sensitivity Tests; D009154:Mutation; D009584:Nitrogen / Q000378:metabolism; D018384:Oxidative Stress; D010641:Phenotype; D013379:Substrate Specificity; D013696:Temperature; D014774:Virulence / Q000235:genetics
D026905:Amino Acid Transport Systems; D000935:Antifungal Agents; D005656:Fungal Proteins; D002244:Carbon; D005947:Glucose; D009584:Nitrogen
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